拟南芥TOC1蛋白特异片段原核表达培养基的优化

通过正交设计实验,优化蛋白诱导培养基中的有机成分和无机成分及各因素之间的相互组合,将各因素对拟南芥生物钟重要组成成分TOC1诱导的影响进行方差分析,发现氧气充足和氧气较缺乏时,培养基最佳组合不同.对各因素进行回归分析得到目的蛋白的培养基最佳配方为:氧气充足时为酵母提取物19g·L-1,NaCl 5g·L-1,DifcoPeptone 25g·L-1 CaCl20.092g·L-1, KCl0.372g·L-1;氧气较缺乏时为酵母提取物19g·L-1,NaCl 5g·L-1,葡萄糖6g·L-1,胰蛋白胨21.7g·L-1 NaH2PO4·2H2O 0.184g·L-1 MgCl20.847g·L-1 KCl 0.372g·L-1.通过进一步的试验发现氧气充足时的最佳配方优于氧气较缺乏时的最佳配方.     

用吸附柱和离交柱层析法从猪血粉水解液中连续分离七种氨基酸

<正> 本文介绍了采用国产活性炭及阳离子交换树脂两种柱体巧妙地组合,并应用不同类型的洗脱剂,变换其浓度和洗脱速度,相互配合,能成功地从猪血粉水解液中一次连续分离和制取L-苯丙氨酸、L-酪氨酸、L-亮氨酸、L-缬氨酸、L-组氨酸、L-赖氨酸和L-精氨酸。多次提取试验表明,工艺操作简单、稳定可靠,苯丙氨酸与酪氨酸、亮氨酸与缬氨酸分离良好,     

亮氨酸和异亮氨酸研究题录选

1　以小什鱼为原料提取亮氨酸的试验 2　猪毛水解物中亮氨酸的离子交换柱分离 3　从猪血水解液中提取亮氨酸 4　以玉米麸质为原料提取L-亮氨酸 5　通过谷氨酸棒状杆菌由α酮异已酸生产L-亮氨酸的微生物生产法 6　L-异亮氨酸生产新技术 7　产L-异亮氨酸新菌株选育及其发酵产物提纯和鉴定 8　生产L-异亮氨酸的新方法 9　异亮氨酸产生菌推理选育思想 10 活细胞反应生产L-异亮氨酸工艺中附产物形成的抑制 11 异亮氨酸生产的新方法 12 L-异亮氨酸产生菌选育的研究 13 L-异亮氨酸质量标准 14 对四种异亮氨质量标准的评价 15 异亮氨酸细菌内毒素和热原检查的探讨 16 从面筋水解物中提取L—亮氨酸 17 从人发中连续提取亮氨酸和胱氨酸工艺初探 　　需要上述资料的读者,请与本刊编辑部联系,编辑部提供复印服务,复印费、邮寄费等共80 元。 　　联系电话：027-87664846 　　邮编：430072 　　地址：武汉市武汉大学《氨基酸和生物资源》编辑部     

营养及环境因素对黄色短杆菌发酵生产L-亮氨酸的影响

目的:研究葡萄糖、玉米浆、蛋氨酸等主要营养物质以及环境因素对发酵生产L-亮氨酸的影响。方法:应用单因素实验确定发酵条件,采用高效液相色谱法测定产量。结果:在最优发酵条件下,以种龄36h、接种量为10%,摇瓶产L-亮氨酸的量可达19.4g/L,采用10L罐分批补料发酵64h可积累29.47g/L的亮氨酸。结论:营养及环境因素对发酵生产L-亮氨酸具有重要影响。     

利用响应面法优化L-组氨酸摇瓶发酵培养基

通过单因素实验对L-组氨酸产生菌LGS4的发酵培养基成分进行筛选,确定了3个影响较大的重要因素,即酵母膏、尿素、硫酸镁。在此基础上,采用二次响应面分析法进行回归分析,得到各因素的最佳水平值。经5批培养验证,预测值与验证试验平均值接近。优化后培养基组成为(g.L-1):蔗糖150,硫酸铵50,酵母膏10,尿素1.2,MgSO42.2,KH2PO41.5,K2HPO40.5,Na2HPO40.5。优化后的发酵培养基使LGS4菌株的L-组氨酸产量提高了15.25%。     

黄色短杆菌TK0303的L-亮氨酸生物合成途径分析

应用途径分析方法分析了在拟稳态时黄色短杆菌(Brevibacterium flavum)TK0303由葡萄糖发酵生产L-亮氨酸的代谢途径,确定了L-亮氨酸合成的最佳途径和最大理论产率。通过比较途径分析所获得的反应模型,确定了丙酮酸和乙酰辅酶A是L-亮氨酸合成途径的关键节点。在此基础上改变外界环境因子,强化L-亮氨酸生物合成途径中丙酮酸和乙酰辅酶A两个关键节点的代谢流,以期进一步提高L-亮氨酸产率。结果表明,经过谷氨酸以及醋酸铵的调节,代谢途径流量发生显著变化,L-亮氨酸产量有明显提高。     

提高红酵母胡萝卜素发酵产率的研究

以红酵母RY-998为实验菌株,采用液体摇瓶发酵方式,在培养基中加入氟化铵等6种添加物,研究它们对红酵母和长及胡萝卜素合成的影响。结果表明;除植酸和二甲基亚砜外,氟化铵,醋酸铵,L-亮氨酸和L-缬氨酸对红酵母胡萝卜素产率均有明显的提高作用;当同时添加氟化铵15mg/L,醋酸铵20mg/L,L-亮氨酸40mg/L和L-缬氨酸30mg/L时,细胞生物量,胡萝卜素含量和产率可分别比对照组提高42.2%,55.4%和121.2%。     

通过谷氨酸棒状杆菌由α-酮异已酸生产L-亮氨酸的微生物生产法

本文研究用谷氨酸棒状杆菌(Co-rlnebacterium glutamicum)将α-酮异巳酸转化为L-亮氨酸的生产过程。在α-酮异已酸的浓度为20克/升的培养基中,经57小时的发酵大约有16克/升的L-亮氨酸被合成,其克分子产量为91%。若采用流加补料分批培养法时,在23小时内就可能从32克/升的α-酮异巳酸中产生出24克升的L-亮氨酸。有关的酶学研究表明,在这种谷氨酸生产菌中a酮异巳酸是经过转氨酶B的作用被转化为亮氨酸的。转氨作用所需的L谷氨酸则通过谷氨酸脱氨酶的作用再生。α-酮异巳酸加入培养基中后,转氨酶B的比活性提高三倍。     

L-亮氨酸的应用及其生产菌的育种思路

氨基酸是构成蛋白质的基本单位,是生物体内不可缺少的营养成份,L-亮氨酸是人体8种必需氨基酸之一,也是3种支链氨基酸之一,在医药、食品、饲料、化妆品等行业具有重要的用途,本文综述了L-亮氨酸的性质、生产方法及发酵法生产的育种思路。     

油樟悬浮培养及次生代谢产物的诱导

以油樟(Cinnamomum longepaniculatum)叶片为外植体在附加6-BA和NAA不同激素浓度组合的MS培养基上筛选质地疏松、生长旺盛的愈伤组织, 分别接种在MS、B5、WPM三种液体培养基中进行细胞悬浮培养, 并检测诱导产生的次生代谢产物。结果表明: 2 mg.L-1 6-BA+ 0.5 mg.L-1 NAA能诱导质地疏松、生长旺盛的愈伤组织, B5基本培养基中细胞长势最好; 愈伤组织在B5+2 mg.L-1 6-BA+ 0.5 mg.L-1 NAA中悬浮培养, 继代2次后形成均一的单细胞; 从油樟悬浮培养物中检测出50%以上的成分是苯甲醇。     

L—亮氨酸发酵条件的研究

对L-亮氨酸产生菌57-4s菌株进行了接种量、装量、pH、发酵时间、碳源、天然营养成份、氨基酸以及生物素、硫酸铵、碳酸钙对L-亮氨酸产量影响的考察,在适宜的条件下,主酸产量高、副酸含量低,L-亮氨酸产量经氨基酸分析仪测定可达24.46mg/ml。     

L-亮氨酸发酵过程氮源反馈与非反馈控制的初步研究

氮源可直接影响氨基酸发酵过程中菌体生长与氨基酸生成。但是关于氨基酸发酵过程中氮源控制的研究报道并不多。本文作者在L-亮氨酸发酵过程碳源流加研究及动力学特征分析基础上,比较了几种非反馈型、反馈型补加硫酸铵的发酵结果,提出了较佳的控制方法。     

Enhancement of bacitracin biosynthesis by branched-chain amino acids in a regulatory mutant ofBacillus licheniformis

DL-4-Azaleucine-resistant mutant of Bacillus licheniformis azlr-1 isolated after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis, was a better bacitracin producer than the parent strain. In the minimal medium, the antibiotic biosynthesis was 4 times higher in the mutant than in the parent strain and less dependent on L-leucine addition. In the complex fermentation medium, the yield was 18-20% higher in the mutant strain. Transaminase B activity measured in the crude extract revealed that the branched-chain amino acid biosynthetic enzymes were 5-10 times derepressed supplying bacitracin synthetase with enhanced quantity of isoleucine and leucine, the building units of bacitracin molecule.     

An Improved Bioprocess for Extracellular l-Leucine Amino Peptidase Production Using Streptomyces gedanensis

A bioprocess was developed for the production of L-leucine aminopeptidase under solid-state fermentation (SSF) by cultivating Streptomyces gedanensis in an inert support impregnated with a minimal medium. Response surface methodology of Box Behnken design was used to derive the optimum level of significant factors (3 ml inoculum (1.2 × 10(9) CFU/ml); 0.275% w/v (NH(4))(2)SO(4); 0.275% w/v MgSO(4)·7H(2)O and 0.55% w/v Tryptone) for maximum LAP production (489 IU/g PUF) as compared to the initial level of 176.3 ± 0.02 IU/g PUF. The high level of extracellular aminopeptidase yield achieved in this work showed the technical feasibility of LAP production under SSF using inert support and is the first report of this kind. The ability of Streptomyces amino peptidase to release particular N-terminal amino acids made them interesting for controlling the degree of hydrolysis and flavor development for a wide range of substrates in food like industries.     

Possible involvement of calcium signaling pathways in L-leucine-stimulated protein synthesis in L6 myotubes

L-Leucine is known to stimulate protein synthesis in L6 myotubes. In the present study, we examined the possible involvement of calcium signaling pathways in the stimulation of protein synthesis induced by L-leucine in L6 myotubes. After 16 h of treatment with L-leucine-depleted medium, the re-addition of L-leucine for 4 h augmented protein synthesis by about 50% as compared with an L-leucine-depleted control. Ryanodine receptor antagonists almost completely abolished the stimulatory effect of L-leucine, while IP(3) receptor antagonists showed partial inhibition when added simultaneously with L-leucine. These results suggest the possibility that calcium signaling pathways are involved in L-leucine-stimulated protein synthesis.     

初始装液比例对赖氨酸产量的影响分析

L-赖氨酸发酵过程中合理且充分供氧是提高发酵产量主要途径之一。大型发酵罐的供氧主要取决搅拌、通气量和装液比例等。通过分析初始装液比例对赖氨酸发酵的影响,发现不同初始装液比例的发酵规律。结果表明,合适的初始装液比例(55%)在相同发酵周期内可以有效提高产量13.54%。     

The effects of phospholipids on the activation of glutamate dehydrogenase by L-leucine.

Glutamate dehydrogenase in disrupted mitochondrial preparations is activated by L-leucine to a much greater extent than is the purified enzyme. A factor, or factors, responsible for modulating the sensitivity of L-leucine is lost during the purification of the enzyme. Although both cardiolipin and phosphatidylserine are inhibitors of the enzyme, only the inhibition by the former phospholipid is reversed by L-leucine. The inhibition of glutamate dehydrogenase by its binding to cardiolipin in the disrupted mitochondrial preparations and its relief by L-leucine could account for the greater sensitivity of such preparations to activation by that amino acid.     

L-leucine auxotrophy in Bifidobacterium globosum

The enzymes involved in the biosynthetic pathway of L-leucine were studied in plasmid-negative and plasmid-positive clones derived from the RU 809 strain of the Bifidobacterium globosum species. The growth of plasmid-positive clones in synthetic medium required L-leucine. We have shown that no detectable activity of the beta-isopropylmalate dehydrogenase enzyme was present in plasmid-positive clones, whereas detectable and significant activity of this enzyme was found in plasmid-negative clones. The lack of activity of the beta-isopropylmalate dehydrogenase enzyme is considered responsible for the L-leucine auxotrophy in the plasmid-positive clones.