产聚γ-谷氨酸菌株的筛选及鉴定

从高温处理过的豆瓣酱等样品中分离获得1株细菌菌株L536,其产物经纸层析分析及氨基酸组成分析鉴定,确定是聚谷氨酸。菌株L536通过形态特征、生理生化特征及16S rRNA基因序列分析,初步鉴定为解淀粉芽孢杆菌(Bacillus amylolique faciens)。该菌株在37℃,180 r/min振荡培养48h,产γ-PGA可达15 g/L。     

High γ-aminobutyric acid production from lactic acid bacteria: Emphasis on Lactobacillus brevis as a functional dairy starter

γ-Aminobutyric acid (GABA) and GABA-rich foods have shown anti-hypertensive and anti-depressant activities as the major functions in humans and animals. Hence, high GABA-producing lactic acid bacteria (LAB) could be used as functional starters for manufacturing novel fermented dairy foods. Glutamic acid decarboxylases (GADs) from LAB are highly conserved at the species level based on the phylogenetic tree of GADs from LAB. Moreover, two functionally distinct GADs and one intact gad operon were observed in all the completely sequenced Lactobacillus brevis strains suggesting its common capability to synthesize GABA. Difficulties and strategies for the manufacture of GABA-rich fermented dairy foods have been discussed and proposed, respectively. In addition, a genetic survey on the sequenced LAB strains demonstrated the absence of cell envelope proteinases in the majority of LAB including Lb. brevis, which diminishes their cell viabilities in milk environments due to their non-proteolytic nature. Thus, several strategies have been proposed to overcome the non-proteolytic nature of Lb. brevis in order to produce GABA-rich dairy foods.     

柱前衍生-高效液相色谱法检测运动型饮料中γ-氨基丁酸的含量

该研究建立了一种测定运动饮料中γ-氨基丁酸（GABA）的柱前衍生-高效液相色谱（HPLC）方法。 结果表明,样品由乙腈萃 取后,上清液经邻苯二甲醛（OPA）于45 ℃条件下在线柱前衍生2.0 min后进行HPLC检测。 色谱条件：Waters sunfire C18为色谱柱,以 20 mmol/L的乙酸钠溶液-乙腈（体积比70∶30）为流动相,二极管阵列检测器（DAD）的波长为247 nm,柱温30 ℃,流速1.0 mL/min, 进样量20 μL,外标法定量。 γ-氨基丁酸在1.0～100.0 μg/mL质量浓度范围内线性关系良好（R2=0.998 6）,检出限为0.30 mg/kg,定量 限为1.0 mg/kg,加标回收率为82.5%～90.6%,相对标准偏差（RSD）为2.9%～3.7%。 采用该方法检测出5种运动型饮品中GABA含量 在0.32～16.38 mg/100 mL范围内。 该方法具有操作简单、专属性强、灵敏度高等优点,可满足运动型饮料中γ-氨基丁酸的检测要求。     

新资源食品——γ-氨基丁酸(GABA)的研究进展

γ-氨基丁酸(GABA)是哺乳动物体内主要的抑制性神经递质,其对人体的功能性和保健作用受到广泛关注。近年来GABA在食品领域中得到较多应用。本文对GABA的功能特性、生产方法及市场应用现状进行综述。     

采用生物转化技术富集大豆制品γ-氨基丁酸研究进展

大豆中含有多种活性成分,具有很高的营养价值及保健功能,开发富含γ-氨基丁酸(GABA)的功能性大豆食品前景广阔。文中综述了大豆富集GABA的方法、途径及采用生物转化技术富集大豆GABA的最新研究进展。     

枯草芽孢杆菌GXA-28发酵生产γ-聚谷氨酸动力学研究

对耐热高产菌株枯草芽孢杆菌GXA-28分批发酵生产γ-聚谷氨酸的动力学特征进行了研究,基于菌体生长特性,结合Logistic方程和Luedeking-Piret方程,提出了菌体生长、产物合成、葡萄糖消耗以及谷氨酸钠消耗的动力学模型。应用Origin8.5对数据进行分析处理,得到了Bacillis subtilis GXA-28分批发酵合成γ-聚谷氨酸相应的动力学参数。将模型预测值和实验值进行比较,结果表明,模型基本反映了枯草芽孢杆菌GXA-28分批发酵过程中的动力学特征。     

Permeabilization of Escherichia coli with ampicillin for a whole cell biocatalyst with enhanced glutamate decarboxylase activity

The activity of whole-cell biocatalysts is strongly compromised by the cell envelope, which is a permeability barrier against the diffusion of substrates and products. Although common chemical or physical permeabilization methods used in cultured cells enhance cell permeability, these methods inevitably add several extra processing steps after cell cultivation, as well as impede large scale processing. To increase membrane permeability and cellbound glutamate decarboxylase(GAD) activity of recombinant Escherichia coli(BL21(DE3)-p ET28a-gad B) cells without the need for an additional permeabilization step, we investigated the permeabilizing effects of adding cell wall synthesis inhibitors or surfactants to the culture media. Ampicillin was the most effective at improving cell-bound GAD activity of the BL21(DE3)-p ET28a-gad B, although it decreased the cell biomass yield. The best permeabilization effect was observed using an ampicillin concentration of 5 μg·ml-1. Using this concentration,the cell biomass did decrease by 40.58%, but the cell-bound GAD activity of BL21(DE3)-p ET28a-gad B and total cell-bound GAD activity per milliliter of culture was enhanced by 6.24- and 3.64-fold, respectively. Treatment of BL21(DE3)-p ET28a-gad B cells with 5 μg·ml-1ampicillin resulted in structural changes to the cell envelope,but did not substantially affect GAD expression. By entrapping the ampicillin-treated cells in an open pore gelation matrix, which is a polymer derived from polyvinyl alcohol(PVA), alginate, and boric acid, the transformation rate of γ-aminobutyric acid(GABA) at the 10 th cycle produced by immobilized and permeabilized cells remained 46% of the first cycle. GAD activity of the immobilized, permeabilized cells remained over 90% after30 days of storage at 4 °C.     

4－氨基－3－苯基丁酸的合成方法

对已知的涉及合成4－氨基－3－苯基丁酸的方法进行了分类、总结,包括涉及硝基苯乙烯的Michael加成反应、以苯基戊二酸为中间体的反应、通过还原氰基或硝基的反应等,从反应原料、反应步骤、产率等方面分析了不同方法的利弊,其中,以糠氯酸为原料的方法最为新颖。为后续工作者在现有成果的基础上进行更深入的研究,寻求更为适用的合成方法提供方便。     

γ-Al2O3表面结构的红外光谱研究

采用原位红外光谱技术对不同温度(100～500℃)脱水γ-Al2O3的表面羟基进行了研究,同时应用吡啶吸附原位红外光谱法对γ-Al2O3表面上路易斯酸(L酸)进行了研究.实验结果表明,在升高脱水温度的过程中,在γ-Al2O3表面共出现五个羟基谱带,分别为3785,3762,3725,3697,3674 cm-1.在γ-Al2O3表面上存在三种L酸性位,分别对应于五配位(酸性最弱)、四配位(酸性中等)和三配位的Al3+(酸性最强).这三种Al3+的数量随脱水温度升高而增大,而且γ-Al2O3表面上的L酸性位均有羟基与其相连.     

以L-谷氨酰胺-锌(II)配合物为供体酶法制备茶氨酸及其反应机理

针对酶法合成茶氨酸中存在的自转肽副反应,合成了L-谷氨酰胺-锌(II)配合物Zn(Gln)2,以其为供体、乙胺为受体、枯草芽孢杆菌B. subtilis GGT为催化剂,酶法制备茶氨酸. 结果表明,Zn(Gln)2在反应主体相稳定性良好,以Zn(Gln)2为供体可有效抑制自转肽副反应;B. subtilis GGT对Zn(Gln)2和Gln的亲和力常数分别为0.53 mmol/L和1.01 mmol/L. 在Zn(Gln)2 6.0 mmol/L、乙胺200 mmol/L及GGT 0.5 U/mL条件下,经37℃反应2 h,茶氨酸浓度最高可达7.38 mmol/L,较以Gln为供体时提高了14.42%.