日本研究人员成功利用大肠杆菌生产聚乳酸

日本丰田汽车、丰田中央研究所和北海道大学的研究小组于2008年10月27日在当天发行的美国科学院院刊（PNAS）早版上发表论文称,利用重组大肠杆菌经生物工艺制造出了含聚乳酸（PLA）的共聚体（copolymer）。此前的做法是先把微生物发酵得到的乳酸转换为环状二聚体丙交酯（lactide）,再通过金属催化剂聚合这种丙交酯,最终合成聚乳酸（丙交酯法）。但是．这种金属催化剂具有毒性,人们一直希望能够确立一种全工序生物工艺聚乳酸合成方法。     

新型乳酸丙交酯成为生物塑料原料

荷兰一家为食品行业生产乳酸的Purac公司日前投资生产含有酒石酸和酒石酸钾钠的乳酸丙交酯,该投资有望为食品、医药和日化工业更廉价有效的生物塑料包装的生产提供原料。新型的丙交酯可以承受175℃的高温,新材料可应用于热罐装瓶,微波加热托盘,耐高温纤维和电子工业等领域。     

可降解生物医用材料面世

由同济大学材料科学与工程学院任杰教授课题组与上海新上化高分子材料有限公司合作承担的“生物可降解材料中间体—DL 丙交酯、L 丙交酯、乙交酯的中试研究”项目日前通过了上海市科委组织的专家鉴定。这些生物可降解材料是以玉米、甘蔗、甜菜等为原料 ,具有良好的生物相容性和生物降解性 ,对人体无毒无刺激 ,且来源丰富、可再生 ,因此成为世界范围的研究开发热点。这些材料在生物医用方面的应用非常广泛。国外聚乳酸、聚乙醇酸在骨科固定、药物控制释放体系、组织工程和医用手术缝合线等方面的应用已有相当的研究、开发 ,并且一些产品已…     

聚丙交酯-乙交酯降解研究进展

本文从分子量、孔径大小和孔径率、力学性能等方面介绍了研究聚丙交酯-乙交酯降解行为的方法,综述了聚丙交酯-乙交酯的化学水解机理和酶催化水解机理,影响聚丙交酯-乙交酯降解速率的内外因素,并比较了聚丙交酯-乙交酯体内外降解的异同,对未来聚丙交酯-乙交酯降解研究的方向提出了展望。     

聚丙交酯- 乙交酯降解研究进展

本文从分子量、孔径大小和孔径率、力学性能等方面介绍了研究聚丙交酯-乙交酯降解行为的方法,综述了聚丙交酯-乙交酯的化学水解机理和酶催化水解机理,影响聚丙交酯-乙交酯降解速率的内外因素,并比较了聚丙交酯-乙交酯体内外降解的异同,对未来聚丙交酯-乙交酯降解研究的方向提出了展望。     

热稳定酯酰辅酶A合成酶的异源表达及酶学特性

【目的】为寻找能合成丙酰辅酶A和丁酰辅酶A等聚酮合成前体的生物催化剂,用体外酶学实验对一个酯酰辅酶A合成酶进行了表征。【方法】利用丙二酰辅酶A合成酶作为输入序列,通过BLAST程序在Caldicellulosiruptor owensensis OL的基因组中找到1个酯酰辅酶A合成酶基因。在大肠杆菌中进行了异源表达,并通过亲和层析进行纯化。底物谱、最适反应条件、稳定性和动力学参数通过体外酶学实验进行表征,而定点突变则用于活性中心的氨基酸残基的分析。【结果】该酶具有较好的底物宽泛性,可识别丙酸、丁酸、2-甲基丙酸、戊酸、3-甲基丁酸、2-甲基丁酸以及环己甲酸等一系列单酸。反应最适温度为30°C,最适p H为7.0。70°C保温8 h后仍有45%的活性残留,表明该酶相对比较稳定。通过活性中心3个位点的定点突变可以改变酶的底物特异性。【结论】C.owensensis OL来源的酯酰辅酶A合成酶是潜在的生物催化剂,可以用于聚酮前体的合成。     

反胶束中单宁酶催化没食子酸烷基酯的生物合成

为了研究单宁酶在有机相中的催化性能,建立了AOT/异辛烷／水反胶束单宁酶催化没食子酸与脂肪醇酯合成反应体系。结果显示：反胶束单宁酶催化体系可成功催化合成C3-C5脂肪醇与没食子酸的酯合成反应。不同反应体系中由于不同脂肪醇的存在,单宁酶的动力学参数和紫外光谱存在差别。结果表明单宁酶对脂肪醇的专一性不强,根据Vmax/Km比值,丁醇与异丁醇是其最适底物,单宁酶催化没食子酸烷基酯合成的动力学符合米氏方程。反应体系中不同的脂肪醇导致了单宁酶构象的差别。     

力复霉素前体甲基丙二酰CoA合成途径的研究

力复霉素合成的碳前体之一(2R)—甲基丙二酰CoA至少可以有三条酶学合成途径。三条途径中的关键酶分别为甲基丙二酰CoA转羧基酶、丙二酰CoA羧化酶、甲基丙二酰CoA变位酶和甲基丙二酰CoA消旋酶。通过比较各个酶活性的时间进程和力复霉素合成时间的相关性,以及各个酶的底物亲合力,对它们在地中海拟无枝酸菌(Amycolatopsis mediterranei)甲基丙二酰CoA合成中的贡献作了排序,发现甲基丙二酰CoA变位酶途径是主要负责酶系。但是各个途径的贡献排序并不是固定不变的,能受到环境因素的调控,丙酸盐的加入将抑制甲基丙二酰CoA变位酶活力,而使得甲基丙二酰CoA转羧基酶成为主要酶系。甲基丙二酰CoA合成途径的多样性有助于细胞对环境变化的灵活反应。此外,对各个酶的调控特性也进行了研究。     

α-氨基酸酯酰基转移酶表达纯化、酶学性质及其催化应用

α-氨基酸酯酰基转移酶(α-amino acid ester acyltransferase,AET)能够催化底物L-丙氨酸甲酯盐酸盐、L-谷氨酰胺合成L-丙氨酰-L-谷氨酰胺(L-alanyl-L-glutamine,丙谷二肽)。利用重组大肠杆菌saet-QC01表达α-氨基酸酯酰基转移酶,对其表达条件进行了优化,通过Ni-NTA亲和层析法分离纯化重组蛋白,并对其酶学性质、催化应用进行了研究。适合酶表达的诱导条件:温度20℃,诱导阶段(OD_(600)=2.0-2.5),IPTG浓度0.6 mmol/L,诱导时间12 h。α-氨基酸酯酰基转移酶的最适反应温度27℃,最适pH 8.5,在pH 7.0-8.0很稳定,在酸性条件下相对稳定,低浓度的Co~(2+)、低浓度的EDTA对酶活有促进作用。在底物浓度丙氨酸甲酯盐酸盐600 mmol/L、谷氨酰胺480 mmol/L,丙谷二肽的产量达到78.2 g/L,生产速率达到1.955 g/(L·min),转化率达到75.0%。α-氨基酸酯酰基转移酶具有良好的酸碱耐受性,催化效率高的优良特性,在工业生产中具有较好的应用潜力。     

环氧基高分子磁性复合微球固定化木瓜蛋白酶的研究

研究以甲基丙烯酸环氧丙酯(GMA)为单体,二甲基丙烯酸乙二醇酯(EDMA)为交联剂,聚乙烯醇(PVA)为分散剂,在Fe3O4磁性纳米粒子存在的条件下,合成了交联度为25%的磁性高分子复合微球(GMAE-DMA).并以这种微球为载体,进行了对木瓜蛋白酶的固定化研究.探讨了最佳的固定化条件如下:温度为25℃,反应时间20h,pH值为8.5,给酶量为160mg/g.同时以酪蛋白为底物,研究了固定化酶的酶学性质,结果表明:固定化酶对不同pH值环境的耐受力、热稳定性和操作稳定性都有较大幅度的提高.实验证明这种高分子磁性复合微球是一种优良的固定化酶载体.     

ENHANCEMENT OF PROPYL GALLATE YIELD IN NONAQUEOUS MEDIUM USING NOVEL CELL-ASSOCIATED TANNASE OF Bacillus massiliensis

Enzymatic synthesis of propyl gallate in organic solvent was studied using cell-associated tannase (EC 3.1.1.20) of Bacillus massiliensis. Lyophilized biomass showing tannase activity was used as the biocatalyst. The effects of solvent, surfactant treatment, and bioimprinting on the propyl gallate synthesis were studied and subsequently optimized. Among various solvents, benzene followed by hexane was found to be the most favorable. Treatment of the biocatalyst with Triton X-100 at a lower concentration (0.2% w/v), before lyophilization, increased the propyl gallate yield by 24.5% compared to the untreated biocatalyst. The biocatalyst was imprinted with various concentrations of gallic acid and tannic acid. Biocatalyst imprinted with tannic acid showed 50% enhancement in the propyl gallate yield compared to the non-imprinted biocatalyst.     

Synthesis and Liposome Formation of New Synthetic Perfluoroalkylated Cationic Lipids Derived from N-[[2-(F-Alkyl)Ethylthio]Methyl (Or Propyl] N'(2-Dimethylaminoethyl) Ureas

Abstract

The synthesis of new urea intermediates was performed from 3′-[2-(F-alkyl)ethylthio]alkyl isocyanates. The quatemization of N-[[2-(F-alkyl)ethyl-thio]methyl (or propyl)] N'-(2-dimethylaminoethyl)ureas by 1-bromoalkanes or 2-(F-hexyl) ethylbromoacetamide allowed to obtain perfluoroalkylated cationic double-chain amphiphiles. Their ability to form liposomes was investigated and discussed according to their molecular structure.     

Selective synthesis of depsipeptides by the immobilized multienzyme enniatin synthetase

Summary The multifunctional enzyme enniatin synthetase was immobilized by adsorption to propyl agarose. The immobilized multienzyme retained 45% of the activity of the free enzyme; an operational half-life of about 15 h was estimated. Selective synthesis of several different enniatin homologues was achieved with propyl agarose-bound enniatin synthetase. In addition to enniatin A, B, and C formation, a selective synthesis of non-naturally occurring depsipeptides, containing norvaline, norleucine, or -aminobutyric acid as sole amino acid moieties, was observed.     

A simple approach to DOTAP and its analogs bearing different fatty acids

A simple synthesis of N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl sulfate (DOTAP) and its analogs differing in the fatty acids is presented. The synthesis is designed as quasi-one-pot reaction and the precipitating products are purified by simple recrystallization.     

Proof of the structure of ristotetraose: synthesis of propyl alpha-ristotetraoside

A definitive synthesis of the propyl glycoside (1) of ristotetraose, the heterotetrasaccharide component of the antibiotic ristomycin A (ristocetin A) is described.     

From lactic acid to poly(lactic acid) (PLA): characterization and analysis of PLA and its precursors

The quality of the monomers lactic acid and lactide as well as the chemical changes induced during polymerization and processing are crucial parameters for controlling the properties of the resulting poly(lactic acid) (PLA) products. This review presents the most important analysis and characterization methods for quality assessment of PLA and its precursors. The impurities typically present in lactic acid or lactide monomers and their possible origins and effects on resulting PLA products are discussed. The significance of the analyses for the different polymer production stages is considered, and special applications of the methods for studying features specific for PLA-based materials are highlighted.     

The effect of parabens on DNA, RNA and protein synthesis in Escherichia coli and Bacillus subtilis

The effect of methyl, propyl and butyl esters of p-hydroxybenzoic acid on DNA and RNA synthesis has been tested in toluenized cells of Escherichia coli and Bacillus subtilis. Both RNA and DNA synthesis of these bacteria were inhibited. The inhibitory concentrations were higher than those previously reported for growth inhibition. Protein synthesis in cell-free extracts (S-30 fraction) of B. subtilis was even more sensitive to parabens than DNA and RNA synthesis, while protein synthesis in Esch. coli was largely unaffected.     

The effect of parabens on DNA, RNA and protein synthesis in Escherichia coli and Bacillus subtilis

The effect of methyl, propyl and butyl esters of p -hydroxybenzoic acid on DNA and RNA synthesis has been tested in toluenized cells of Escherichia coli and Bacillus subtilis. Both RNA and DNA synthesis of these bacteria were inhibited. The inhibitory concentrations were higher than those previously reported for growth inhibition. Protein synthesis in cell-free extracts (S-30 fraction) of B. subtilis was even more sensitive to parabens than DNA and RNA synthesis, while protein synthesis in Esch. coli was largely unaffected.     

Continuous production of kyotorphin

The continuous alpha-chymotrypsin-catalyzed peptide synthesis of kyotorphin, tyrosyl-arginine, via the N(alpha) formyltyrosyl-arginine propyl ester is described. For continuous process development, two reaction systems were studied: immobilized alpha-chymotrypsin covalently bound to Eupergit C packed in a column, and soluble alpha-chymotrypsin utilizing an enzyme membrane reactor. Selectivities and kinetic parameters are discussed. The use of soluble enzyme in an enzyme membrane reactor proved superior to the covalently immobilized enzyme. A significant loss of enzyme activity and a certain decrease of selectivity was observed during immobilization. It was shown that the addition of organic solvent, in this case n-propanol, causes a severe diminuation of the enzyme activity.     

The interaction of α-chymotrypsin with isosteric substrates of different charge type

1. The synthesis of three substrates of alpha-chymotrypsin of closely similar steric requirements but different charge type is reported. 2. The interaction of these compounds [SS-dimethyl-(l-3-carboxymethyl-3-acetamido)propyl sulphonium iodide, l-2-acetamido-5-methylhexanoic acid methyl ester and N-acetyl-l-glutamic acid alpha-methyl ester] with alpha-chymotrypsin has been studied. 3. For the charged substrates, values of k(0) are two orders of magnitude smaller than, and values of K(m) two orders of magnitude larger than, the corresponding values for the uncharged isostere. 4. The results are interpreted in terms of the known specificity of the enzyme, and the relationship between binding and kinetic specificities is discussed.