This work was funded by a grant from the National Institutes of Health (R15 AI117670) to B.M.S and P.J.W, as well as a Cottrell College Scholar Award from the Research Corporation (20185) to P.J.W. L.M.M. was supported by a Provost's Fellowship from CMU.

<ol>
	<li>Elbein, A. D., Pan, Y. T., Pastuszak, I., Carroll, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+insights+on+trehalose:+a+multifunctional+molecule.">New insights on trehalose: a multifunctional molecule.</a> Glycobiology. 13, 17-27 (2003).</li><li>Tournu, H., Fiori, A., Van Dijck, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relevance+of+trehalose+in+pathogenicity:+some+general+rules,+yet+many+exceptions.">Relevance of trehalose in pathogenicity: some general rules, yet many exceptions.</a> PLoS Pathog. 9, 1003447 (2013).</li><li>Ohtake, S., Wang, Y. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trehalose:+Current+use+and+future+applications.">Trehalose: Current use and future applications.</a> J. Pharm. Sci. 100, 2020-2053 (2011).</li><li>Adams, R. P., Kendall, E., Kartha, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+free+sugars+in+growing+and+desiccated+plants+of+Selaginella+lepidophylla.">Comparison of free sugars in growing and desiccated plants of Selaginella lepidophylla.</a> Biochem. Syst. Ecol. 18, 107-110 (1990).</li><li>Kubota, M., Ohnishi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Glycoenzymes. , (2000).</li><li>Walmagh, M., Zhao, R., Desmet, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trehalose+analogues:+latest+insights+in+properties+and+biocatalytic+production.">Trehalose analogues: latest insights in properties and biocatalytic production.</a> Int. J. Mol. Sci. 16, 13729-13745 (2015).</li><li>Kim, H. -. M., Chang, Y. -. K., Ryu, S. -. I., Moon, S. -. G., Lee, S. -. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enzymatic+synthesis+of+a+galactose-containing+trehalose+analogue+disaccharide+by+Pyrococcus+horikoshii+trehalose-synthesizing+glycosyltransferase:+Inhibitory+effects+on+several+disaccharidase+activities.">Enzymatic synthesis of a galactose-containing trehalose analogue disaccharide by Pyrococcus horikoshii trehalose-synthesizing glycosyltransferase: Inhibitory effects on several disaccharidase activities.</a> J. Mol. Catal. B: Enzym. 49, 98-103 (2007).</li><li>Backus, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake+of+unnatural+trehalose+analogs+as+a+reporter+for+Mycobacterium+tuberculosis.">Uptake of unnatural trehalose analogs as a reporter for Mycobacterium tuberculosis.</a> Nat. Chem. Biol. 7, 228-235 (2011).</li><li>Swarts, B. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probing+the+mycobacterial+trehalome+with+bioorthogonal+chemistry.">Probing the mycobacterial trehalome with bioorthogonal chemistry.</a> J. Am. Chem. Soc. 134, 16123-16126 (2012).</li><li>Rose, J. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+biological+evaluation+of+trehalose+analogs+as+potential+inhibitors+of+mycobacterial+cell+wall+biosynthesis.">Synthesis and biological evaluation of trehalose analogs as potential inhibitors of mycobacterial cell wall biosynthesis.</a> Carbohydr. Res. 337, 105-120 (2002).</li><li>Wang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+trehalose-based+compounds+and+their+inhibitory+activities+against+Mycobacterium+smegmatis.">Synthesis of trehalose-based compounds and their inhibitory activities against Mycobacterium smegmatis.</a> Bioorg. Med. Chem. 12, 6397-6413 (2004).</li><li>Gobec, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design,+synthesis,+biochemical+evaluation+and+antimycobacterial+action+of+phosphonate+inhibitors+of+antigen+85C,+a+crucial+enzyme+involved+in+biosynthesis+of+the+mycobacterial+cell+wall.">Design, synthesis, biochemical evaluation and antimycobacterial action of phosphonate inhibitors of antigen 85C, a crucial enzyme involved in biosynthesis of the mycobacterial cell wall.</a> Eur. J. Med. Chem. 42, 54-63 (2007).</li><li>Sarpe, V. A., Kulkarni, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regioselective+protection+and+functionalization+of+trehalose.">Regioselective protection and functionalization of trehalose.</a> Trends in Carbohydr. Res. 5, 8-33 (2013).</li><li>Chaube, M. A., Kulkarni, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stereoselective+construction+of+1,1-alpha,alpha-glycosidic+bonds.">Stereoselective construction of 1,1-alpha,alpha-glycosidic bonds.</a> Trends in Carbohydr. Res. 4, 1-19 (2013).</li><li>Leigh, C. D., Bertozzi, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthetic+studies+toward+Mycobacterium+tuberculosis+sulfolipid-I.">Synthetic studies toward Mycobacterium tuberculosis sulfolipid-I.</a> J. Org. Chem. 73, 1008-1017 (2008).</li><li>Chaen, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+enzymatic+synthesis+of+disaccharide,+alpha-D-galactosyl-alpha-D-glucoside,+by+trehalose+phosphorylase+from+Thermoanaerobacter+brockii.">Efficient enzymatic synthesis of disaccharide, alpha-D-galactosyl-alpha-D-glucoside, by trehalose phosphorylase from Thermoanaerobacter brockii.</a> J. Appl. Glycosci. 48, 135-137 (2001).</li><li>Vander Borght, J., Soetaert, W., Desmet, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+the+acceptor+specificity+of+trehalose+phosphorylase+for+the+production+of+trehalose+analogs.">Engineering the acceptor specificity of trehalose phosphorylase for the production of trehalose analogs.</a> Biotechnol. Progr. 28, 1257-1262 (2012).</li><li>Kouril, T., Zaparty, M., Marrero, J., Brinkmann, H., Siebers, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+trehalose+synthesizing+pathway+in+the+hyperthermophilic+Crenarchaeon+Thermoproteus+tenax:+the+unidirectional+TreT+pathway.">A novel trehalose synthesizing pathway in the hyperthermophilic Crenarchaeon Thermoproteus tenax: the unidirectional TreT pathway.</a> Arch. Microbiol. 190, 355-369 (2008).</li><li>Urbanek, B. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemoenzymatic+synthesis+of+trehalose+analogues:+rapid+access+to+chemical+probes+for+investigating+mycobacteria.">Chemoenzymatic synthesis of trehalose analogues: rapid access to chemical probes for investigating mycobacteria.</a> ChemBioChem. 15, 2066-2070 (2014).</li><li>Rostovtsev, V. V., Green, L. G., Fokin, V. V., Sharpless, K. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+stepwise+Huisgen+cycloaddition+process:+copper(I)-catalyzed+regioselective+"ligation"+of+azides+and+terminal+alkynes.">A stepwise Huisgen cycloaddition process: copper(I)-catalyzed regioselective "ligation" of azides and terminal alkynes.</a> Angew. Chem. Int. Ed. 41, 2596-2599 (2002).</li><li>Torn&#248;e, C. W., Christensen, C., Meldal, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peptidotriazoles+on+solid+phase:+[1,2,3]-triazoles+by+regiospecific+copper(I)-catalyzed+1,3-dipolar+cycloadditions+of+terminal+alkynes+to+azides.">Peptidotriazoles on solid phase: [1,2,3]-triazoles by regiospecific copper(I)-catalyzed 1,3-dipolar cycloadditions of terminal alkynes to azides.</a> J. Org. Chem. 67, 3057-3064 (2002).</li><li>Kalscheuer, R., Weinrick, B., Veeraraghavan, U., Besra, G. S., Jacobs, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trehalose-recycling+ABC+transporter+LpqY-SugA-SugB-SugC+is+essential+for+virulence+of+Mycobacterium+tuberculosis.">Trehalose-recycling ABC transporter LpqY-SugA-SugB-SugC is essential for virulence of Mycobacterium tuberculosis.</a> Proc. Natl. Acad. Sci. U. S. A. 107, 21761-21766 (2010).</li></ol>

The authors have nothing to disclose.

トレハロース類似体は、食品や医薬品の保全から微生物感染症6の診断と治療のために、さまざまな分野に影響を与える可能性を持っています。既存の多段階の化学合成法（ 例えば 、天然に複雑なマイコバクテリア糖を生じる）の変形例の複数の部位を有する複雑なトレハロース類似体を製造するために有用です。しかしながら、これらの方法は、比較的簡単な一置?...

トレハロースは、1,1-α、αグリコシド結合（ 図1A）によって結合された2つのグルコース部分からなる対称な非還元性二糖類です。トレハロースは、ヒトおよび他の哺乳動物には存在しないが、それは、細菌、真菌、植物、無脊椎動物1に一般的に見出されます。ほとんどの生物におけるトレハロースの主な役割は、乾燥1などの環境ストレスから保護することです。また、いくつかのヒト病原体は、細胞エンベロープ生合成のメディエーターとしておよび免疫脂質2の構築のためのビルディングブロックとしてトレハロースを用いる結核を引き起こす結核菌を含む、毒性のためにトレハロースを必要とします。 <img alt="図1" src="/files/ftp_upload/54485/54485fig1.jpg" /> 図1： トレハロースとトレハロース類似体。 （Xは構造的な変更であり、天然トレハロースと不自然なトレハロースアナログの）構造。 （B）バイオプリザベーションとバイオイメージングにおける潜在的なアプリケーションを持っている文献に報告トレハロースアナログの例を。 、そのユニークな構造と生理機能に、トレハロースは、バイオ（テクノ）で使用するための論理的および生物医学的応用3を大きな注目を集めています。 nature- 例えばで観察されたトレハロースの保護特性は、その印象的な能力は4 -haveはバイオプリザベーション・アプリケーションでその広範な使用に拍車をかけ極度の脱水を受けた&quot;復活&quot;植物の生活を維持するのに役立ちます。トレハロースは、核酸、タンパク質、細胞、および組織3のような生物学的サンプルの広い配列を保存するために使用されてきました。例えば、トレハロースは、医薬品tの数で安定化添加剤として使用されています帽子は3いくつかの抗癌モノクローナル抗体を含む、市販されています。同様に、トレハロースは、食品業界で甘味料として使用され、それが広く両方食品および化粧品産業における製品の保存のために使用されます。商用アプリケーションのこれらの種類のトレハロースの採用は、当初、天然の供給源から、または合成により純粋なトレハロースの大量を得ることができないことによって制限されていました。しかしながら、澱粉からトレハロースの経済的な生産のための効率的な酵素的方法は、最近、広範な商業的使用に拍車をかけている、開発されています 5。 （ 図1Aに示される一般構造、 図1（b）に示すトレハロース類似体の具体例）化学トレハロース類似体と呼ぶ、トレハロース誘導体を改変、様々な用途にますます注目を集めている6。例えば、ラクトトレハロース、ガラクトースで置き換え、そのグルコース単位のいずれかのトレハロースアナログであり、従って、その4位のヒドロキシル基が反転立体化学構成を有しています。ラクトトレハロースはトレハロースと同様の安定化特性を有するが、ノンカロリー食品添加物6,7などが魅力的、腸の酵素による分解に耐性です。 トレハロース類似体における当社グループの持分は、主にマイコバクテリア特異的プローブおよび阻害剤としてのそれらの値に関係します。バリーとデイビス基は蛍光顕微鏡8によりその検出を可能にする、生結核菌の細胞壁を標識代謝することが示されたFITCケトトレハロース命名フルオレセイン共役ケトトレハロースアナログを開発しました。ベルトッツィラボは、代謝、細胞壁にラベルを付け、その後DET可能性が小さいアジドトレハロース（TreAz）類似体を開発しましたクリック化学と蛍光分析9を使用してected。これらの進歩は、結核のための画像診断剤としてトレハロースベースのプローブを使用する可能性を指します。トレハロース類似体はまた、生存率および病原10、11、12のために必須である細菌で経路を破壊するために、それらの潜在的に結核菌の阻害剤として追求されてきました。 これまで、バイオ（テクノ）論理及び生物医学的用途のためにトレハロースアナログを開発する主な障害は、効率的な合成方法がないことです。トレハロースの類似体を製造するには2つの伝統的なルートは、化学合成（ 図2）に依存しています。他が適切に官能単糖ビルディングブロックで始まり、に化学的グリコシル化を実行する必要が、一方の経路は、天然トレハロースの非対称化/変更を伴います1,1-α、αグリコシド結合を築きます。最近の総説13、14で議論されてきたこれらのアプローチは、 結核菌 15からそのようなスルホ脂質-1のような複雑なトレハロース含有天然物、少量の多段階合成を達成するために有用であることが判明しました。しかしながら、両方のアプローチは、非化学者に一般的に、時間がかかり、非効率的なアクセス不能であり、そして、さらに、環境に優しいとはみなされません。したがって、トレハロース類似体の特定の種類を合成するため、これらの戦略は理想的ではありません。 <img alt="図2" src="/files/ftp_upload/54485/54485fig2.jpg" /> 図2： トレハロースアナログ合成へのアプローチ。化学は難しいPROTECを伴う多段階の手順を使用し、左に示し、トレハロースアナログ合成へのアプローチ化/脱保護、非対称化、および/またはグリコシル化のステップ。右側に示す酵素合成は、立体選択的に水溶液中で類似体をトレハロースするための簡単な、保護されていない基質を変換する酵素（複数可）を使用しています。本明細書で報告酵素プロトコルは、単一のステップでトレハロースの類似体へのグルコース類似体およびUDP-グルコースを変換するトレハロース合成酵素（TRET）酵素を使用しています。 <a href="http://ecsource.jove.com/files/ftp_upload/54485/54485fig2large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> トレハロースアナログへの効率的な生体触媒ルートは、製造、評価し、分子のこの有望なクラスの適用を容易にするであろう。トレハロースの生産5用の市販の酵素処理は、それが基質として澱粉を使用しているため類縁体を合成に適応可能ではありませんが、他の生合成経路がありますトレハロースアナログ合成のために利用することができる自然の中での方法。しかし、最近6を検討したこの地域での研究は、制限されています。一つのレポートには、対応するフルオログルコースからの単一フルオロトレハロースアナログにアクセスするには、 大腸菌（Escherichia coli）トレハロース生合成経路に触発されたメソッドを使用していました。しかしながら、このアプローチは、限られた効率と汎用8を有する三酵素系を必要とします。探索された別のアプローチは、原理的にはグルコースアナログおよびグルコース-1-リン酸6、16、17からトレハロース類似体の一段階合成を可能にする逆方向にトレハロースホス（TREP）を使用することです。このアプローチは、将来有望であることができるが、両方の反転と保持TrePs現在アナログ合成のための欠点を有します。例えば、反転TrePsは法外EXPEを持っていますnsiveドナー分子（β-D-グルコース1-リン酸）と保持TrePsが悪い酵素発現収量/安定性と限られた基板の混乱を持っています。 TREP媒介アナログ合成が実用的である前に（酵素工学を介して、例えば ）重要な改善が必要とされるであろう。 現時点では、トレハロース類似体の酵素合成のための最も実用的なアプローチは、グルコースおよびウリジン二リン酸（UDP）は、単一のステップ6でトレハロースに-グルコース変換トレハロース合成酵素（TRET）酵素を使用することです。我々は最近、グルコース類似体およびUDP-グルコース（ 図3）19からトレハロース類似体を合成-to サーモテナックス TRET、耐熱性および一方向の酵素18の使用を報告しました。この酵素は、合成方向に動作し、TREPのシステムで検出されたトレハロースの劣化の問題を回避できます。このワンステップ反応coul1時間で完了さD、及び（高速液体クロマトグラフィー（HPLC）によって決定されるように&gt; 99％まで）トレハロースアナログの広範な種々の高収率でアクセスされた容易に入手可能なグルコース類似体基質から（代表的な結果の表1を参照セクション）。 <img alt="図3" src="/files/ftp_upload/54485/54485fig3.jpg" /> 図3： トレハロースアナログのTRET触媒による一段階合成。 T.テナックスからTRET酵素は、立体選択的に1ステップでトレハロースの類似体を形成するために容易に利用可能なグルコース類似体およびUDP-グルコースに参加することができます。 R 1〜R 4 =可変構造変更、例えばアジド、フルオロ、デオキシ、チオ、立体化学、または同位体ラベルの変更のため、 Yは、例えば、酸素または硫黄、または同位体標識されたヘテロための変数のヘテロ原子を=。 ここでは、広告を提供します発現および精製TRETの大腸菌から、最適化されたTRET反応条件、および完全に水性相で行われる改良された精製法を含む、TRET合成処理するためのプロトコルをetailed。この修正されたプロトコルは、セミ分取スケール（10-100 mg）を上の多様なトレハロースアナログの好都合かつ効率的な合成及び精製を可能にします。我々はまた、マイコバクテリア細胞の迅速な蛍光検出を有効にして1時間未満でマイコバクテリアにトレハロースベースのプローブを調製し、投与するために、このプロトコルの使用を実証します。

<table><tbody><tr><td>LB agar</td><td>Research Products International</td><td>L24021</td><td></td></tr><tr><td>Ampicillin sodium salt</td><td>Sigma Aldrich</td><td>A9518</td><td></td></tr><tr><td>Luria broth</td><td>Research Products International</td><td>L24045</td><td></td></tr><tr><td>Terrific Broth</td><td>Research Products International</td><td>T15050</td><td></td></tr><tr><td>L-(+)-Arabinose</td><td>Sigma Aldrich</td><td>A3256</td><td></td></tr><tr><td>Phosphate-buffered Saline</td><td>GE Healthcare</td><td>SH30256</td><td></td></tr><tr><td>Imidazole</td><td>Sigma Aldrich</td><td>I5513</td><td></td></tr><tr><td>Sodium chloride</td><td>BDH</td><td>BDH9286</td><td></td></tr><tr><td>Sodium phosphate, monobasic</td><td>Fisher Scientific</td><td>S374</td><td></td></tr><tr><td>Syringe filter, 0.45 &amp;micro;m</td><td>Fisher Scientific</td><td>09719D</td><td></td></tr><tr><td>Protease Inhibitor mini-tablets, EDTA-free</td><td>Thermo Scientific</td><td>88666</td><td></td></tr><tr><td>HisTrap HP nickel affinity column, 5 mL</td><td>GE Healthcare</td><td>17-5248-02</td><td></td></tr><tr><td>TRIS base ultrapure</td><td>Research Products International</td><td>T60040</td><td></td></tr><tr><td>Dialysis tubing, MWCO 12&amp;ndash;14,000</td><td>Fisher Scientific</td><td>21-152-16</td><td></td></tr><tr><td>Glucose analogues</td><td>CarboSynth, Sigma Aldrich, Santa Cruz Biotechnology, American Radiolabeled Chemicals</td><td></td><td>Examples of vendors that offer numerous glucose analogues</td></tr><tr><td>6-Azido-6-deoxy glucopyranose (6-GlcAz)</td><td>CarboSynth</td><td>MA02620</td><td></td></tr><tr><td>UDP-Glucose</td><td>abcam Biochemicals</td><td>ab120384</td><td></td></tr><tr><td>Magnesium chloride hexahydrate&amp;nbsp;</td><td>Fisher Scientific</td><td>M33</td><td></td></tr><tr><td>Amicon Ultra-15 centrifugal filter unit</td><td>EMD Millipore</td><td>UFC901008</td><td></td></tr><tr><td>Bio-Rex RG 501-X8 mixed-bed ion-exchange resin</td><td>Bio-Rad</td><td>444-9999</td><td></td></tr><tr><td>Extra-Fine Bio-Gel P2 media</td><td>Bio-Rad</td><td>150-4118</td><td></td></tr><tr><td>Glass-backed silica gel thin-layer chromatography plates</td><td>EMD Millipore</td><td>1056280001</td><td></td></tr><tr><td>&lt;em&gt;n&lt;/em&gt;-Butanol</td><td>Fisher Scientific</td><td>A399</td><td></td></tr><tr><td>Ethanol</td><td>Fisher Scientific</td><td>S25310A</td><td></td></tr><tr><td>Sulfuric acid</td><td>Fisher Scientific</td><td>A300</td><td></td></tr><tr><td>Acetonitrile</td><td>EMD Millipore</td><td>AX0145</td><td></td></tr><tr><td>Deuterium oxide, 99.8%</td><td>Acros Organics</td><td>351430075</td><td></td></tr><tr><td>Aminopropyl HPLC column</td><td>Sigma Aldrich</td><td>58338</td><td></td></tr><tr><td>Bovine serum albumin</td><td>Sigma Aldrich</td><td>5470</td><td></td></tr><tr><td>Para-formaldehyde</td><td>Ted Pella</td><td>18505</td><td></td></tr><tr><td>Alkyne-488</td><td>Sigma Aldrich</td><td>761621</td><td></td></tr><tr><td>Sodium ascorbate</td><td>Sigma Aldrich</td><td>A7631</td><td></td></tr><tr><td>Tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA)</td><td>Click Chemistry Tools</td><td>1061</td><td></td></tr><tr><td>&lt;em&gt;tert&lt;/em&gt;-Butanol</td><td>Sigma Aldrich</td><td>360538</td><td></td></tr><tr><td>Dimethylsulfoxide</td><td>Sigma Aldrich</td><td>W387520</td><td></td></tr><tr><td>Copper(II) sulfate</td><td>Sigma Aldrich</td><td>C1297</td><td></td></tr><tr><td>Fluoromount-G mounting medium</td><td>Southern Biotechnology</td><td>10001</td><td></td></tr></tbody></table>

rapid one-step enzymatic synthesis and all-aqueous purification of trehalose analogues

化学的に修飾されたトレハロースのバージョン、またはトレハロース類似体は、他の分野の中で、生物学、バイオテクノロジー、および製薬科学の用途を有します。例えば、検出可能なタグを有するトレハロースアナログは結核菌を検出するために使用されており、結核診断用造影剤としての用途を有することができます。トレハロースの加水分解的に安定したバージョンはまた、ノンカロリー甘味料およびbioprotective剤としての使用のためのそれらの潜在的に追求されています。様々なアプリケーションのための化合物のこのクラスの魅力にもかかわらず、彼らの可能性は、それらの製造のための強固なルートがないために満たされていないままです。ここでは、化学合成に関連する問題を回避トレハロース類似体の迅速かつ効率的な一段階の生体触媒合成のための詳細なプロトコルを報告します。 サーモテナックスから耐熱性トレハロースシンターゼ（TRET）酵素を利用することにより、トレハロースアナログはgeneratすることができグルコース類似体および15から60分で（定量的変換まで）高い収率でウリジン二リン酸グルコースから一工程で編。スピン透析とイオン交換で構成されて簡単かつ迅速な非クロマトグラフィー精製プロトコルは、わずか45として分で水溶液中の濃度が既知の多くのトレハロースの類似体を提供することができます。未反応のグルコース類似体が残る場合には、トレハロースアナログ生成物のクロマトグラフィー精製を行うことができます。全体的に、この方法は効率的で、非化学者にアクセス可能であるトレハロース類似体の迅速合成および精製のための「グリーン」生体触媒のプラットフォームを提供します。この方法の適用を例示するために、我々は、合成、すべての水精製、および1時間未満とマイコバクテリアの有効蛍光検出を取ったそのすべてがマイコバクテリアのトレハロースベースのクリック化学プローブの投与のためのプロトコルについて説明します。今後は、OTHの中で、ということを想定しますERアプリケーションは、このプロトコルは、結核の診断のためのトレハロースベースのプローブの迅速な合成に適用することができます。例えば、短寿命放射性核種修飾トレハロースアナログ（ 例えば 、18 F修飾トレハロース）は、陽電子放出断層撮影、コンピュータ断層撮影法（PET-CT）などの高度な臨床イメージング様式を使用することができます。

T.テナックス TRETは、標準的なタンパク質発現および精製技術を使用して、約4ミリグラム/ Lの収量で大腸菌から得ました。単一ニッケルアフィニティークロマトグラフィー工程は、（代表FPLCのトレースを図4に示されている）、大腸菌溶解物からTRETを精製するのに十分でした。 TRET合成プロセスに私たちの最初の出版物に設立されたよ...

Watch this Scientific Journal Video about Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues at JoVE.com

Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues

Trehalose analogues are emerging as important molecules for bio(techno)logical and biomedical applications. We describe an optimized protocol for enzymatically synthesizing and purifying trehalose analogues that is simple, efficient, fast, and environmentally friendly. Its application to the rapid production and administration of a probe for the detection of mycobacteria is demonstrated.

迅速なワンステップ酵素合成とトレハロース類似体のすべての水精製

Chemically modified versions of trehalose, or trehalose analogues, have applications in biology, biotechnology, and pharmaceutical science, among other ...

rapid-one-step-enzymatic-synthesis-all-aqueous-purification-trehalose

Research

JoVE Journal

Biochemistry

10.3K ビュー.  Central Michigan University. Trehalose analogues are emerging as important molecules for bio(techno)logical and biomedical applications. We describe an optimized protocol for enzymatically synthesizing and purifying trehalose analogues that is simple, efficient, fast, and environmentally friendly. Its application to the rapid production and administration of a probe for the detection of mycobacteria is demonstrated.

ビデオ: Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

The traditional strategy for the introduction of chemical functionalities is the use of solid-phase synthesis by appending suitably modified phosphoramidite precursors to the nascent chain. However, the conditions used during the synthesis and the restriction to rather short sequences hamper the applicability of this methodology. On the other hand, modified nucleoside triphosphates are activated building blocks that have been employed for the mild introduction of numerous functional groups into nucleic acids, a strategy that paves the way for the use of modified nucleic acids in a wide-ranging palette of practical applications such as functional tagging and generation of ribozymes and DNAzymes. One of the major challenges resides in the intricacy of the methodology leading to the isolation and characterization of these nucleoside analogues.
	
	In this video article, we present a detailed protocol for the synthesis of these modified analogues using phosphorous(III)-based reagents. In addition, the procedure for their biochemical characterization is divulged, with a special emphasis on primer extension reactions and TdT tailing polymerization. This detailed protocol will be of use for the crafting of modified dNTPs and their further use in chemical biology.

The protocol described herein aims to explain and abridge the numerous obstacles in the way of the intricate route leading to modified nucleoside triphosphates. Consequently, this protocol facilitates both the synthesis of these activated building-blocks and their availability for practical applications.

ヌクレオシド三リン酸 - 合成からの生化学的特徴付けへ

The  traditional strategy for the introduction of chemical functionalities is the  use of solid-phase synthesis by appending suitably modified ...

化学

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design efficacious vaccines carriers. Nanoparticle-based platforms can prolong the persistence of vaccine antigens, which could improve vaccine immunogenicity1. Several biodegradable polymers have been studied as vaccine delivery vehicles1; in particular, polyanhydride particles have demonstrated the ability to provide sustained release of stable protein antigens and to activate antigen presenting cells and modulate immune responses2-12.
The molecular design of these vaccine carriers needs to integrate the rational selection of polymer properties as well as the incorporation of appropriate targeting agents. High throughput automated fabrication of targeting ligands and functionalized particles is a powerful tool that will enhance the ability to study a wide range of properties and will lead to the design of reproducible vaccine delivery devices.
The addition of targeting ligands capable of being recognized by specific receptors on immune cells has been shown to modulate and tailor immune responses10,11,13 C-type lectin receptors (CLRs) are pattern recognition receptors (PRRs) that recognize carbohydrates present on the surface of pathogens. The stimulation of immune cells via CLRs allows for enhanced internalization of antigen and subsequent presentation for further T cell activation14,15. Therefore, carbohydrate molecules play an important role in the study of immune responses; however, the use of these biomolecules often suffers from the lack of availability of structurally well-defined and pure carbohydrates. An automation platform based on iterative solution-phase reactions can enable rapid and controlled synthesis of these synthetically challenging molecules using significantly lower building block quantities than traditional solid-phase methods16,17.
Herein we report a protocol for the automated solution-phase synthesis of oligosaccharides such as mannose-based targeting ligands with fluorous solid-phase extraction for intermediate purification. After development of automated methods to make the carbohydrate-based targeting agent, we describe methods for their attachment on the surface of polyanhydride nanoparticles employing an automated robotic set up operated by LabVIEW as previously described10. Surface functionalization with carbohydrates has shown efficacy in targeting CLRs10,11 and increasing the throughput of the fabrication method to unearth the complexities associated with a multi-parametric system will be of great value (Figure 1a).

In this article, a high throughput method is presented for the synthesis of oligosaccharides and their attachment to the surface of polyanhydride nanoparticles for further use in targeting specific receptors on antigen presenting cells.

無水物ナノ粒子のハイスループット糖鎖の合成と機能化

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design ...

生物工学

Chemo-enzymatic Synthesis of N-glycans for Array Development and HIV Antibody Profiling

We present a highly efficient way for the rapid preparation of a wide range of N-linked oligosaccharides (estimated to exceed 20,000 structures) that are commonly found on human glycoproteins. To achieve the desired structural diversity, the strategy began with the chemo-enzymatic synthesis of three kinds of oligosaccharyl fluoride modules, followed by their stepwise &#945;-selective glycosylations at the 3-O and 6-O positions of the mannose residue of the common core trisaccharide having a crucial &#946;-mannoside linkage. We further attached the N-glycans to the surface of an aluminum oxide-coated glass (ACG) slide to create a covalent mixed array for the analysis of hetero-ligand interaction with an HIV antibody. In particular, the binding behavior of a newly isolated HIV-1 broadly neutralizing antibody (bNAb), PG9, to the mixture of closely spaced Man5GlcNAc2 (Man5) and 2,6-di-sialylated bi-antennary complex type N-glycan (SCT) on an ACG array, opens a new avenue to guide the effective immunogen design for HIV vaccine development. In addition, our ACG array embodies a powerful tool to study other HIV antibodies for hetero-ligand binding behavior.

Chemo-enzymatic Synthesis of N-glycans for Array Development and HIV Antibody Profiling

A modular approach to the synthesis of N-glycans for attachment to an aluminum oxide-coated glass slide (ACG slide) as a glycan microarray has been developed and its use for the profiling of an HIV broadly neutralizing antibody has been demonstrated.

N型糖鎖アレイ開発および HIV 抗体のための化学酵素合成プロファイリング

We present a highly efficient way for the rapid preparation of a wide range of N-linked oligosaccharides (estimated to exceed 20,000 structures) that are ...

One-pot Microwave-assisted Conversion of Anomeric Nitrate-esters to Trichloroacetimidates

The goal of the following procedure is to provide a demonstration of the one-pot conversion of a 2-azido-1-nitrate-ester to a trichloroacetimidate glycosyl donor. Following azido-nitration of a glycal, the product 2-azido-1-nitrate ester can be hydrolyzed under microwave-assisted irradiation. This transformation is usually achieved using strongly nucleophilic reagents and extended reaction times. Microwave irradiation induces hydrolysis, in the absence of reagents, with short reaction times. Following denitration, the intermediate anomeric alcohol is converted, in the same pot, to the corresponding 2-azido-1-trichloroacetimidate.

A 2-azido-1-nitrate-ester can be converted to the corresponding 2-azido-1-trichloroacetimidate in a one-pot procedure. The goal of the manuscript is to demonstrate utility of the microwave reactor in carbohydrate synthesis.

1 つの鍋マイクロ波への変換アノマー硝酸エステルの Trichloroacetimidates

The goal of the following procedure is to provide a demonstration of the one-pot conversion of a 2-azido-1-nitrate-ester to a trichloroacetimidate ...

Constructing Thioether/Vinyl Sulfide-tethered Helical Peptides Via Photo-induced Thiol-ene/yne Hydrothiolation

Here, we describe a detailed protocol for the preparation of thioether-tethered peptides using on-resin intramolecular/intermolecular thiol-ene hydrothiolation. In addition, this protocol describes the preparation of vinyl-sulfide-tethered peptides using in-solution intramolecular thiol-yne hydrothiolation between amino acids that possess alkene/alkyne side chains and cysteine residues at i, i+4 positions. Linear peptides were synthesized using a standard Fmoc-based solid-phase peptide synthesis (SPPS). Thiol-ene hydrothiolation is carried out using either an intramolecular thio-ene reaction or an intermolecular thio-ene reaction, depending on the peptide length. In this research, an intramolecular thio-ene reaction is carried out in the case of shorter peptides using on-resin deprotection of the trityl groups of cysteine residues following the complete synthesis of the linear peptide. The resin is then set to UV irradiation using photoinitiator 4-methoxyacetophenone (MAP) and 2-hydroxy-1-[4-(2-hydroxyethoxy)-phenyl]-2-methyl-1-propanone (MMP). The intermolecular thiol-ene reaction is carried out by dissolving Fmoc-Cys-OH in an N,N-dimethylformamide (DMF) solvent. This is then reacted with the peptide using the alkene-bearing residue on resin. After that, the macrolactamization is carried out using benzotriazole-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBop), 1-hydroxybenzotriazole (HoBt), and 4-Methylmorpholine (NMM) as activation reagents on the resin. Following the macrolactamization, the peptide synthesis is continued using standard SPPS. In the case of the thio-yne hydrothiolation, the linear peptide is cleaved from the resin, dried, and subsequently dissolved in degassed DMF. This is then irradiated using UV light with photoinitiator 2,2-dimethoxy-2-phenylacetophenone (DMPA). Following the reaction, DMF is evaporated and the crude residue is precipitated and purified using high-performance liquid chromatography (HPLC). These methods could function to simplify the generation of thioether-tethered cyclic peptides due to the use of the thio-ene/yne click chemistry that possesses superior functional group tolerance and good yield. The introduction of thioether bonds into peptides takes advantage of the nucleophilic nature of cysteine residues and is redox-inert relative to disulfide bonds.

We present a protocol for the construction of thioether/vinyl sulfide-tethered helical peptides using photo-induced thiol-ene/thiol-yne hydrothiolation.

チオエーテル/ビニール硫化テザー ヘリカル ペプチドによる光誘起チオール-エン/イン Hydrothiolation を構築します。

Here, we describe a detailed protocol for the preparation of thioether-tethered peptides using on-resin intramolecular/intermolecular thiol-ene ...

Regioselective O-Glycosylation of Nucleosides via the Temporary 2',3'-Diol Protection by a Boronic Ester for the Synthesis of Disaccharide Nucleosides

Disaccharide nucleosides, which consist of disaccharide and nucleobase moieties, have been known as a valuable group of natural products having multifarious bioactivities. Although chemical O-glycosylation is a commonly beneficial strategy to synthesize disaccharide nucleosides, the preparation of substrates such as glycosyl donors and acceptors requires&#160;tedious protecting&#160;group manipulations and a purification at each synthetic step. Meanwhile, several research groups have reported that boronic and borinic esters serve as a protecting or activating group of carbohydrate derivatives to achieve the regio- and/or stereoselective acylation, alkylation, silylation, and glycosylation. In this article, we demonstrate the procedure for the regioselective O-glycosylation of unprotected ribonucleosides utilizing boronic acid. The esterification of 2&#39;,3&#39;-diol of ribonucleosides with boronic acid makes the temporary protection of diol, and, following O-glycosylation with a glycosyl donor in the presence of p-toluenesulfenyl chloride and silver triflate, permits the regioselective reaction of the 5&#39;-hydroxyl group to afford the disaccharide nucleosides. This method could be applied to various nucleosides, such as guanosine, adenosine, cytidine, uridine, 5-metyluridine, and 5-fluorouridine. This article and the accompanying video represent&#160;useful (visual) information for the O-glycosylation of unprotected nucleosides and their analogs for the synthesis of not only disaccharide nucleosides, but also a variety of biologically relevant derivatives.

Regioselective O-Glycosylation of Nucleosides via the Temporary 2',3'-Diol Protection by a Boronic Ester for the Synthesis of Disaccharide Nucleosides

Here, we present protocols for the synthesis of disaccharide nucleosides by the regioselective O-glycosylation of ribonucleosides via a temporary protection of their 2&#39;,3&#39;-diol moieties utilizing a cyclic boronic ester. This method applies to several unprotected nucleosides such as adenosine, guanosine, cytidine, uridine, 5-methyluridine, and 5-fluorouridine to give corresponding disaccharide nucleosides.

ヌクレオシドを介して一時 2'、3'-ジオール保護ホウ酸エステル糖ヌクレオシドの合成のためのOの位置選択的グリコシル化反応

Disaccharide nucleosides, which consist of disaccharide and nucleobase moieties, have been known as a valuable group of natural products having ...

Hierarchical and Programmable One-Pot Oligosaccharide Synthesis

This article presents a general experimental protocol for programmable one-pot oligosaccharide synthesis and demonstrates how to use Auto-CHO software for generating potential synthetic solutions. The programmable one-pot oligosaccharide synthesis approach is designed to empower fast oligosaccharide synthesis of large amounts using thioglycoside building blocks (BBLs) with the appropriate sequential order of relative reactivity values (RRVs). Auto-CHO is a cross-platform software with a graphical user interface that provides possible synthetic solutions for programmable one-pot oligosaccharide synthesis by searching a BBL library (containing about 150 validated and &#62;50,000 virtual BBLs) with accurately predicted RRVs by support vector regression. The algorithm for hierarchical one-pot synthesis has been implemented in Auto-CHO and uses fragments generated by one-pot reactions as new BBLs. In addition, Auto-CHO allows users to give feedback for virtual BBLs to keep valuable ones for further use. One-pot synthesis of stage-specific embryonic antigen 4 (SSEA-4), which is a pluripotent human embryonic stem cell marker, is demonstrated in this work.

This protocol demonstrates how to use the Auto-CHO software for hierarchical and programmable one-pot synthesis of oligosaccharides. It also describes the general procedure for RRV determination&#160;experiments and one-pot glycosylation of SSEA-4.

階層的でプログラム可能なワンポットオリゴ糖合成

This article presents a general experimental protocol for programmable one-pot oligosaccharide synthesis and demonstrates how to use Auto-CHO software for ...

Versatile CO2 Transformations into Complex Products: A One-pot Two-step Strategy

CO2 transformations using a one-pot two-step method are presented herein. The purpose of the method is to give access to a variety of value-added products and notably to generate chiral carbon centers. The crucial first step consists in the selective double hydroboration of CO2 catalyzed by an iron hydride complex. The product obtained with this 4 e- reduction is a rare bis(boryl)acetal, compound 1, which is subjected in situ to three different reactions in a second step. The first reaction concerns a condensation reaction with (diisopropyl)phenylamine affording the corresponding imine 2. In the second and third reaction, intermediate 1 reacts with triazol-5-ylidene (Enders&#39; carbene) to afford compounds 3 or 4, depending on the reaction conditions. In both compounds, C-C bonds are formed, and chiral centers are generated from CO2 as the only source of carbon. Compound 4 exhibits two chiral centers obtained in a diastereoselective manner in a formose-type mechanism. We proved that the remaining boryl fragment plays a key role in this unprecedented stereocontrol. The interest of the method stands on the reactive and versatile nature of 1, giving rise to various complex molecules from a single intermediate. The complexity of a two-step method is compensated by the overall short reaction time (2 h for the larger reaction time), and mild reaction conditions (25 &#176;C to 80 &#176;C and 1 to 3 atm of CO2).

Versatile CO2 Transformations into Complex Products: A One-pot Two-step Strategy

CO2 transformations are conducted in a one-pot two-step procedure for the synthesis of complex molecules. The selective 4 e- reduction of CO2 with a hydroborane reductant affords a reactive and versatile bis(boryl)acetal intermediate which is subsequently involved in condensation reaction or carbene-mediated C-C coupling generation.

多彩なCO2の複雑な製品への変革:ワンポット2段階戦略

CO2 transformations using a one-pot two-step method are presented herein. The purpose of the method is to give access to a variety of value-added products ...

Synthesis of Information-bearing Peptoids and their Sequence-directed Dynamic Covalent Self-assembly

This protocol presents the use of Lewis acidic multi-role reagents to circumvent kinetic trapping observed during the self-assembly of information-encoded oligomeric strands mediated by paired dynamic covalent interactions in a manner mimicking the thermal cycling commonly employed for the self-assembly of complementary nucleic acid sequences. Primary amine monomers bearing aldehyde and amine pendant moieties are functionalized with orthogonal protecting groups for use as dynamic covalent reactant pairs. Using a modified automated peptide synthesizer, the primary amine monomers are encoded into oligo(peptoid) strands through solid-phase submonomer synthesis. Upon purification by high-performance liquid chromatography (HPLC) and characterization by electrospray ionization mass spectrometry (ESI-MS), sequence-specific oligomers are subjected to high-loading of a Lewis acidic rare-earth metal triflate which both deprotects the aldehyde moieties and affects the reactant pair equilibrium such that strands completely dissociate. Subsequently, a fraction of the Lewis acid is extracted, enabling annealing of complementary sequence-specific strands to form information-encoded molecular ladders characterized by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). The simple procedure outlined in this report circumvents kinetic traps commonly experienced in the field of dynamic covalent assembly and serves as a platform for the future design of robust, complex architectures.

A protocol is presented for the synthesis of information-encoded peptoid oligomers and for the sequence-directed self-assembly of these peptoids into molecular ladders using amines and aldehydes as dynamic covalent reactant pairs and Lewis acidic rare-earth metal triflates as multi-role reagents.

情報を持つペプトイドの合成と、その配列指向動的共有自己集合

This protocol presents the use of Lewis acidic multi-role reagents to circumvent kinetic trapping observed during the self-assembly of information-encoded ...

Assembly and Operation of an Acoustofluidic Device for Enhanced Delivery of Molecular Compounds to Cells

Efficient intracellular delivery of biomolecules is required for a broad range of biomedical research and cell-based therapeutic applications. Ultrasound-mediated sonoporation is an emerging technique for rapid intracellular delivery of biomolecules. Sonoporation occurs when cavitation of gas-filled microbubbles forms transient pores in nearby cell membranes, which enables rapid uptake of biomolecules from the surrounding fluid. Current techniques for in vitro sonoporation of cells in suspension are limited by slow throughput, variability in the ultrasound exposure conditions for each cell, and high cost. To address these limitations, a low-cost acoustofluidic device has been developed which integrates an ultrasound transducer in a PDMS-based fluidic device to induce consistent sonoporation of cells as they flow through the channels in combination with ultrasound contrast agents. The device is fabricated using standard photolithography techniques to produce the PDMS-based fluidic chip. An ultrasound piezo disk transducer is attached to the device and driven by a microcontroller. The assembly can be integrated inside a 3D-printed case for added protection. Cells and microbubbles are pushed through the device using a syringe pump or a peristaltic pump connected to PVC tubing. Enhanced delivery of biomolecules to human T cells and lung cancer cells is demonstrated with this acoustofluidic system. Compared to bulk treatment approaches, this acoustofluidic system increases throughput and reduces variability, which can improve cell processing methods for biomedical research applications and manufacturing of cell-based therapeutics.

This protocol describes the assembly and operation of a low-cost acoustofluidic device for rapid molecular delivery to cells via sonoporation induced by ultrasound contrast agents.

迅速なワンステップ酵素合成とトレハロース類似体のすべての水精製

要約

さらに動画を探す

この動画の章

ヌクレオシド三リン酸 - 合成からの生化学的特徴付けへ

無水物ナノ粒子のハイスループット糖鎖の合成と機能化

N型糖鎖アレイ開発および HIV 抗体のための化学酵素合成プロファイリング

1 つの鍋マイクロ波への変換アノマー硝酸エステルの Trichloroacetimidates

チオエーテル/ビニール硫化テザーヘリカルペプチドによる光誘起チオール-エン/イン Hydrothiolation を構築します。

ヌクレオシドを介して一時 2'、3'-ジオール保護ホウ酸エステル糖ヌクレオシドの合成のためのOの位置選択的グリコシル化反応

階層的でプログラム可能なワンポットオリゴ糖合成

多彩_なCO2の複雑な製品への変革:ワンポット2段階戦略

情報を持つペプトイドの合成と、その配列指向動的共有自己集合

細胞への分子化合物の送達を強化するためのアキュトー流体デバイスの組立と操作

迅速なワンステップ酵素合成とトレハロース類似体のすべての水精製

要約

さらに動画を探す

この動画の章

ヌクレオシド三リン酸 - 合成からの生化学的特徴付けへ

無水物ナノ粒子のハイスループット糖鎖の合成と機能化

N型糖鎖アレイ開発および HIV 抗体のための化学酵素合成プロファイリング

1 つの鍋マイクロ波への変換アノマー硝酸エステルの Trichloroacetimidates

チオエーテル/ビニール硫化テザー ヘリカル ペプチドによる光誘起チオール-エン/イン Hydrothiolation を構築します。

ヌクレオシドを介して一時 2'、3'-ジオール保護ホウ酸エステル糖ヌクレオシドの合成のためのOの位置選択的グリコシル化反応

階層的でプログラム可能なワンポットオリゴ糖合成

多彩なCO2の複雑な製品への変革:ワンポット2段階戦略

情報を持つペプトイドの合成と、その配列指向動的共有自己集合

細胞への分子化合物の送達を強化するためのアキュトー流体デバイスの組立と操作

チオエーテル/ビニール硫化テザーヘリカルペプチドによる光誘起チオール-エン/イン Hydrothiolation を構築します。

多彩_なCO2の複雑な製品への変革:ワンポット2段階戦略