促进剂对辅酶Q10生物合成的影响

目的研究不同的添加物对辅酶Q10发酵合成的影响。方法辅酶Q10用皂化法提取,高效液相法检测。结果添加与辅酶Q10的合成有关物质如对羟基苯甲酸、胡萝卜汁、西红柿汁及烟叶,均能提高辅酶Q10的产量,分别提高6.3%、9.0%、18.1%、12.1%。     

Catalysis of an Essential Step in Vitamin B2 Biosynthesis by a Consortium of Broad Spectrum Hydrolases

An enzyme catalysing the essential dephosphorylation of the riboflavin precursor, 5‐amino‐6‐ribitylamino‐2,4(1H,3H)‐pyrimidinedione 5′‐phosphate ( 6 ), was purified about 800‐fold from a riboflavin‐producing Bacillus subtilis strain, and was assigned as the translation product of the ycsE gene by mass spectrometry. YcsE is a member of the large haloacid dehalogenase (HAD) superfamily. The recombinant protein was expressed in Escherichia coli. It catalyses the hydrolysis of 6 (v_max, 12 μmol mg^?1 min^?1; K_M, 54 μm ) and of FMN (v_max, 25 μmol mg^?1 min^?1; K_M, 135 μm ). A ycsE deletion mutant of B. subtilis was not riboflavin dependent. Two additional proteins (YwtE, YitU) that catalyse the hydrolysis of 6 at appreciable rates were identified by screening 13 putative HAD superfamily members from B. subtilis. The evolutionary processes that have resulted in the handling of an essential step in the biosynthesis of an essential cofactor by a consortium of promiscuous enzymes require further analysis.     

脱氮假单胞杆菌耗氧合成V_(B_(12))发酵条件的研究

对脱氮假单胞杆菌(Pseudomonas denitri ficans)耗氧合成V_(B_(12))的培养模式研究表明,分批补料发酵比分批发酵合成V_(B_(12))提高71.42%;其中液糖补料与糖蜜补料在同等条件下,产物浓度前者比后者高出19.08%。最适V_(B_(12))合成的培养条件如种龄、接种量和初始pH值分别为18h,10%和7.0;发酵过程中摇床的转速对V_(B_(12))的合成影响显著,其中最佳转速为260r/min。在以上优化条件下,V_(B_(12))的最高单位能达到140 g/L。     

Detailed Structure–Function Correlations of Bacillus subtilis Acetolactate Synthase

Isobutanol is deemed to be a next‐generation biofuel and a renewable platform chemical. 1 Non‐natural biosynthetic pathways for isobutanol production have been implemented in cell‐based and in vitro systems with Bacillus subtilis acetolactate synthase (AlsS) as key biocatalyst. 2 – 6 AlsS catalyzes the condensation of two pyruvate molecules to acetolactate with thiamine diphosphate and Mg²⁺ as cofactors. AlsS also catalyzes the conversion of 2‐ketoisovalerate into isobutyraldehyde, the immediate precursor of isobutanol. Our phylogenetic analysis suggests that the ALS enzyme family forms a distinct subgroup of ThDP‐dependent enzymes. To unravel catalytically relevant structure‐function relationships, we solved the AlsS crystal structure at 2.3 Å in the presence of ThDP, Mg²⁺ and in a transition state with a 2‐lactyl moiety bound to ThDP. We supplemented our structural data by point mutations in the active site to identify catalytically important residues.     

Chloroplastic glycolipids fuel aldehyde biosynthesis in the marine diatom Thalassiosira rotula

Enzymatic preparations and specialized analytical tools have shown that chloroplast-derived glycolipids are the main substrates for the biosynthetic pathway that produces antiproliferative polyunsaturated aldehydes in broken cells of the marine diatom Thalassiosira rotula. This process, which is associated with the formation of free fatty acids and lyso compounds from polar lipids but not triglycerides, is largely dependent on glycolipid hydrolytic activity, rather than phospholipase A(2) as previously suggested. Preliminary characterization of lipolytic enzymes has revealed protein bands of 40-45 kDa. Under native conditions these proteins seem to be associated with soluble aggregates that have an apparent molecular weight of approximately 200 kDa. The biochemical process, which is similar to that described in the algal-bloom forming diatom Skeletonema costatum, suggests a mechanism based on decompartmentalization and mixing of preexisting enzymes and substrates.     

紫杉醇及其类似物生产的代谢工程研究进展

大量生产抗癌新药紫杉醇是目前生物技术的研究热点之一。通过对紫杉醇生物合成途径中关键酶基因的分离、克隆与遗传转化等方面最新研究结果进行评述 ,指出通过紫杉醇次生代谢途径中关键酶的基因改造及遗传转化 ,构建高效表达紫杉醇或其重要前体紫杉烷的基因工程细胞株 ,并建立其相应的基因表达调控方法 ,是解决紫杉醇药源问题的最有前途的方法之一。     

BIOTECHNOLOGY AND BIOENGINEERING Roles of biomolecules in the biosynthesis of silver nanoparticles: Case of gardenia jasminoides extract

Rapid development of biosynthesis of metal nanoparticles using plants has attracted extensive interests to further investigate this novel and eco-friendly method. In the biosynthesis process, the plant may act as reducing agent, capping agent or shape directing agent. However, identifying specific roles of various components in the plant is challenging. In this study, we use biosynthesis of silver nanoparticles with Gardenia jasminoides Ellis ex-tract to address this issue. The formation process of silver nanoparticles is investigated and the nanoparticles are characterized with the ultraviolet-visible spectroscopy, Fourier transform infrared spectra and scanning electron mi-croscopy. The results indicate that the Gardenia jasminoides Ellis extract can reduce silver ions to form silver nanoparticles, stabilize the nanoparticles, and affect the growth of silver nanocrystal to form silver nanowires. Only geniposide in the extract exhibits good shape-directing ability for silver nanowires. It is found that bovine albumin is a potential capping agent, whereas rutin, gallic acid and chlorogenic acid possess reducing and capping ability.     

根霉LN-1生物合成天麻素条件研究

以对羟基苯甲醛为底物,探索根霉LN-1生物合成天麻素的最适条件。采用静息细胞转化法,通过分析转化时间、温度、pH值、底物质量浓度等不同因素对转化率的影响,获得了根霉LN-1的最适转化条件为:12 g(湿质量)静息细胞/120 mL转化液,底物质量浓度2.3 mg/mL,pH=5.5,28℃转化24 h。在此基础上,转化液经萃取、硅胶柱色谱分离得到转化产物,通过理化性质和波谱分析鉴定其结构。经HPLC、NMR、EI-MS等测定,证明所得转化产物为天麻素。利用微生物的还原和糖基化作用一步转化合成天麻素,该方法的建立为天麻素的合成开辟了一条新的途径。