生物合成γ-氨基丁酸的乳酸菌的筛选

γ-氨基丁酸(GABA)具有降血压、提高脑活力、改善更年期综合症等生理活性,GABA的生物合成主要采用大肠杆菌为菌种。通过筛选微生物,尝试利用乳酸菌生物合成γ-氨基丁酸,得到一株食品安全(GRAS)乳酸菌SYFS1.009,利用该菌株表现出较高的合成GABA活性,通过发酵技术可以获得较高含量的γ-氨基丁酸产品。     

高产γ-氨基丁酸乳酸菌菌株的诱变选育与鉴定

本文以从内蒙古传统发酵食品中分离的80株乳酸菌为研究对象,在GYP培养基中进行高产γ-氨基丁酸(GABA)菌株的筛选后,利用紫外线进行诱变处理,得到GABA突变菌株,并对其进行了菌种鉴定。结果表明,从80株供试乳酸菌中筛选出4株高产γ-氨基丁酸的菌株,再经紫外诱变后得到1株高产突变菌株US3-3。该菌株紫外诱变后,其γ-氨基丁酸含量为2.482 g/L,是诱变前提高1.9倍,并对其多次传代稳定性较好,经16S r DNA序列分析,鉴定为乳酸片球菌(Pediococcus acidilactici)。     

高产γ-氨基丁酸霉菌菌株的筛选及诱变育种

以霉菌作为出发菌株,将其接入大豆-水(3:5,m/V)的发酵培养基中进行发酵,根据发酵过程中γ-氨基丁酸(GABA)产量,筛选出GABA的高产霉菌菌株,然后利用紫外线对高产菌株进行诱变处理,并通过抗性初筛获得长势较好的突变菌株,再分别利用含有质量浓度为1g/100mL L-谷氨酸的PDA综合培养基和大豆水溶液的发酵培养基进行两次筛选,得到稳定高产GABA突变菌株。结果表明,通过对产GABA霉菌菌株的筛选,得到米曲霉3.800为高产GABA霉菌,GABA产量达到0.674g/L。以米曲霉3.800作为原始菌株,对其进行紫外诱变处理,从而获得高产GABA突变菌株,结果表明,利用紫外线对米曲霉3.800进行处理的最佳照射时间为2min。在此照射时间下,通过突变菌株在含有质量浓度为1g/100mL L-谷氨酸的PDA综合培养基和大豆-水溶液的发酵培养基进行两次筛选,最终获得一株稳定的高产GABA突变菌株3.800-4,其在含有质量浓度为1g/100mL谷氨酸的PDA综合培养基中的GABA产量为4.491g/L,比原菌株的GABA产量提高了23.58%;同时其在大豆-水的发酵培养基中的GABA产量为0.874g/L,比原菌株的产量提高了29.67%。     

产γ-氨基丁酸乳酸菌的分离鉴定及发酵条件优化

从四川泡菜水中分离筛选出一株产γ-氨基丁酸(GABA)能力较强的乳酸菌W1-9,根据菌落和个体形态、生理生化指标及16S r DNA序列的系统鉴定,菌株W1-9为植物乳杆菌(Lactobacillus plantarum)。菌株W1-9在含10 g/L谷氨酸(Glu)的MRS培养基中培养3 d,可产生2.18 g/L GABA。通过对菌株发酵培养基及发酵条件优化,结果表明:以黄瓜汁为发酵培养基,初始p H 5.5,底物谷氨酸钠(MSG)添加量12 g/L,菌液接种量1.2%,在该条件下GABA产量达到7.62 g/L。     

水牛乳中高产γ-氨基丁酸乳酸菌的筛选与鉴定

从新鲜水牛乳中筛选出三株高产γ-氨基丁酸(GABA)的菌株G-24、G-25、G-29。通过菌株形态特征、生理生化特征和16S r DNA序列分析,鉴定这三株菌均为乳酸乳球菌乳酸亚种(Lactococcus lactis subsp.lactis),将其分别接种于10 m L含0.01 g/m L谷氨酸的MRS液体培养基中,37℃培养72 h,其产γ-氨基丁酸含量均可达60 mg/100 m L以上。     

产γ-氨基丁酸乳酸菌的分离筛选

从土壤、泡菜、酸奶等样品中分离、筛选出产γ-氨基丁酸(GABA)的乳酸菌,获得较高产GABA的乳酸菌6#菌株,利用HPLC-ELSD检测法精确测定出乳酸菌6#发酵样品中GABA的含量达到0.463g/L,并对此菌株进行了初步的鉴定.     

GABA高产菌株的筛选、鉴定及诱变选育

从火龙果果实表面上筛选出一株发酵产γ-氨基丁酸(GABA)白色菌株,经形态学观察、生理生化试验和18S rDNA测序分析,鉴定为假丝酵母菌菌株(Candida.sp),命名为C2。C2作为出发菌株,分别采用紫外线(UV)和亚硝基胍(NTG)诱变方法选育高产γ-氨基丁酸菌株。与出发菌株相比,紫外诱变菌株γ-氨基丁酸产量增加了40.25%,亚硝基胍诱变菌株γ-氨基丁酸产量增加了62.83%。通过紫外线和亚硝基胍复合诱变,得到正向突变株,其中Y6突变株遗传性状稳定,γ-氨基丁酸产量达2.561 g/L,产量比诱变前提高了3.1倍。     

寒地黑土中产γ-氨基丁酸乳酸菌筛选及发酵培养基优化

利用MRS培养基分别从牡丹江、漠河、五常的寒地黑土中筛选产γ-氨基丁酸(GABA)的乳酸菌菌株,对其做形态学观察和生理生化鉴定,并进行了发酵培养基组成的优化实验,分析比较不同碳源、氮源、碳氮比、初始pH和L-谷氨酸浓度对产GABA的影响。结果表明:共筛出五株菌株,分别编号为SbO001、SbO002、SbO003、SbO004、SbO005,可以将五株菌株初步鉴定为链球菌科(Streptococcaceae)。当发酵培养基组分碳源为葡萄糖,氮源为硫酸铵,碳氮比1:1,初始pH为5.5,底物为1%的L-谷氨酸时,菌株产GABA的能力较好,为1.112 g/L。本实验将为寒地黑土中产GABA乳酸菌的筛选提供一定的理论依据。     

产γ-氨基丁酸乳酸菌的分离筛选及其主要性能

γ-氨基丁酸(GABA)是一种天然存在的功能性氨基酸,具有降血压、改善脑功能、镇静、增强长期记忆及提高肝、肾机能等功效.可利用微生物发酵制得GABA.本文从西藏干酪、乳酸饮料、自制泡菜等样品中分离到7株乳酸菌,通过HPLC-AFS法检测GABA含量,筛选出3株GABA高产乳酸菌株,再用GYP培养基30℃恒温厌氧培养48 h后,其GABA产量分别达到383、345和813 μg/mL.通过对产量最高的ML7菌株进行菌落、菌体形态观察和API20A系统生理、生化特性鉴定,初步确定其为发酵乳杆菌(Lactobacillus fermentum).     

泡菜中1株高产γ-氨基丁酸且具有抗菌性乳酸菌株的筛选和鉴定

利用薄层层析法从泡菜乳酸菌中筛选出1株高产γ-氨基丁酸(γ-aminobutyric acid,GABA)的菌株,通过API50CHL糖发酵测定、16S r DNA序列分析鉴定该菌为短乳杆菌,并命名为短乳杆菌(Lactobacillus brevis)PD1-2。经高效液相色谱法定量测定,该菌在含10 g/L谷氨酸钠(sodium glutamate,MSG)的MRS培养基中发酵48 h,生产GABA的转化率达100%;在含50 g/L MSG的MRS培养基中发酵72 h,GABA的转化率达93.7%。通过抗菌性实验发现,该菌对革兰氏阳性菌的藤黄微球菌、枯草芽孢杆菌、蜡状芽孢杆菌、单增李斯特菌和革兰氏阴性菌的大肠杆菌均具有抗菌性,且初步确定抑菌物质为细菌素。     

Determination of ethanol,volatile fatty acids,lactic and succinic acids in fermentation liquids by gas chromatography

A rapid and simple quantitative method was developed to determine, by gas chromatography, the concentrations in fermentation liquids of ethanol, the C₂-C₆ volatile fatty acids, and lactic and succinic acids. Aqueous samples were acidified with 250μlml^?1 metaphosphoric acid (5:1 ratio), centrifuged, and injected directly on to a column containing a porous aromatic polymer (Chromosorb 101) maintained at 200°C in a gas chromatograph fitted with a flame ionisation detector. It was unnecessary to purify samples further before injection, although distillation and ion-exchange methods were examined. Derivatisation of lactic and succinic acids before injection was not necessary, but the lowest level of detection of these two relatively non-volatile acids was about four times greater than that for the volatile fatty acids. The method described was suitable for the analysis of rumen fluid, methane digester fluid, silage extracts and other anaerobic fermentation fluids. The relative retention times are given for 23 organic acids and six other fermentation end-products.     

Effects of abomasal infusions of fatty acids and 1-carbon donors on apparent fatty acid digestibility and incorporation into milk fat in cows

Our primary objective was to determine the effects of the abomasal infusion of 16-carbon (16C) and 22-carbon (22C) fatty acids (FA) on apparent FA digestibility, plasma FA concentrations, and their incorporation into milk fat in cows. Our secondary objective was to study the effects of 1-carbon donors choline and l-serine on these variables. Five rumen-cannulated Holstein cows (214 ± 4.9 d in milk; 3.2 ± 1.1 parity) were enrolled in a 5 × 5 Latin square experiment with experimental periods lasting 6 d. Abomasal infusates consisted of (1) palmitic acid (PA; 98% 16:0 of total fat), (2) PA + choline chloride (PA+CC; 50 g/d of choline chloride), (3) PA + l-serine (PA+S; 170 g/d of l-serine), (4) behenic acid (BA; 92% 22:0 of total fat), and (5) docosahexaenoic acid algal oil (DHA; 47.5% DHA of total fat). Emulsions were formulated to provide 301 g/d of total FA and were balanced to provide a minimum of 40 and 19 g/d of 16:0 and glycerol, respectively, to match the content found in the infused algal oil. Apparent digestibility of FA was highest in DHA, intermediate in PA, and lowest in BA. Digestibility of 16C FA was lowest in BA and highest in PA. The digestibility of 22C FA was highest in DHA relative to BA (99 vs. 58%), whereas 1-carbon donors had no effect on 22C FA digestibility. Plasma 16C FA concentrations were greatest with PA treatment, and 22C FA concentrations were ~3-fold greater in DHA-treated cows relative to all other treatments. Milk fat 16:0 content was highest in PA relative to BA and DHA (e.g., 37 vs. 27% in PA and DHA), whereas the milk yield of 16:0 was higher in PA relative to DHA (i.e., 454 vs. 235 g/d). Similarly, milk 22:0 content and yield were ~10-fold higher in BA relative to all other treatments, whereas DHA treatment resulted in higher content and yield of 22:6 in milk fat relative to all other treatments (41- and 38-fold higher, respectively). Consequently, the content of FA >16C (i.e., preformed) was higher in milk fat from cows infused with BA and DHA relative to PA. De novo FA content in milk did not differ between PA, PA+CC, and PA+S (~16% of milk fat) but was higher in BA and DHA treatments (19 and 21%, respectively). We conclude that FA carbon chain length and degree of saturation affected FA digestibility and availability for absorption as well as their incorporation into milk fat. The abomasal infusion of choline chloride and l-serine did not modify these variables relative to infusing palmitic acid alone.     

Short communication: Eicosatrienoic acid and docosatrienoic acid do not promote vaccenic acid accumulation in mixed ruminal cultures

Previous research found that docosahexaenoic acid (C22:6n-3) was a component of fish oil that promotes trans-C18:1 accumulation in ruminal cultures when incubated with linoleic acid. The objective of this study was to determine if eicosatrienoic acid (C20:3n-3) and docosatrienoic acid (C22:3n-3), n-3 fatty acids in fish oil, promote accumulation of trans-C18:1, vaccenic acid (VA) in particular, using cultures of mixed ruminal microorganisms. Treatments consisted of control, control plus 5 mg of C20:3n-3 (ETA), control plus 5 mg of C22:3n-3 (DTA), control plus 15 mg of linoleic acid (LA), control plus 5 mg of C20:3n-3 and 15 mg of linoleic acid (ETALA), and control plus 5 mg of C22:3n-3 and 15 mg of linoleic acid (DTALA). Treatments were incubated in triplicate in 125-mL flasks, and 5 mL of culture contents was taken at 0 and 24 h for fatty acid analysis by gas-liquid chromatography. After 24 h of incubation, the concentrations of trans-C18:1 (0.87, 0.88, and 0.99 mg/culture), and VA (0.52, 0.56, and 0.62 mg/culture) were similar for the control, ETA, and DTA cultures, respectively. The concentrations of trans-C18:1 (5.51, 5.41, and 5.36 mg/culture), and VA (4.78, 4.62, and 4.59 mg/culture) were also similar between LA, ETALA, and DTALA cultures, respectively. These data suggest that C20:3n-3 and C22:3n-3 are not the active components in fish oil that promote VA accumulation when incubated with linoleic acid.     

奶粉脂肪酸与乳制品风味关系研究

用气质（GC—MS）联用色谱分析了11个商业奶粉样品的脂肪酸组成以及含量,每个样品均检测到了28种脂肪酸,在表现奶粉风味的4个呈味脂肪酸,也即辛酸、己酸、壬酸和葵酸中只检测到了辛酸和葵酸。辛酸和葵酸含量在进口奶粉中普遍高于国产奶粉。国产奶粉中辛酸和葵酸的含量以2号最好,3号其次。亚油酸含量在国产奶粉中普遍高于进口奶粉。     

气相色谱法同时快速测定食品中丙酸、山梨酸、苯甲酸及脱氢乙酸

通过液液萃取净化样品研究,建立了食品中丙酸、山梨酸、苯甲酸、脱氢乙酸及其盐含量气相色谱同时快速测定方法,适用于固体非酯(脂)类食品的检测。结果表明:丙酸的回收率在85.1%~91.3%之间,其余3种防腐剂的回收率均在95.2%~99.4%之间;实验室内变异系数(CV,n=6)最大值≤4.7%,4种防腐剂检出限均在0.002 g/kg以下。4种目标物在有杂质干扰时,可用不同的极性毛细管柱做进一步的确认。本方法具有适用范围广、检测效率高、重现性好、准确度高、检出限低的特点,推广应用对我国食品安全的监督检验具有重要的意义。     

花生四烯酸、二十二碳六烯酸和二十碳五烯酸 在炎症中的作用概述

心脑血管疾病、肿瘤、糖尿病、神经系统疾病、自身免疫等疾病严重危害着人类的生命和健康,并消耗着大量医疗资源。事实上,很多疾病发生和发展的背后都伴随着炎症反应,炎症是众多疾病的病理基础,甚至是导致这些疾病的诱因。炎症本身是机体的防御性反应,但过度的炎症反应和长期慢性炎症会损害机体的稳态。炎症的调节和控制由炎症介质介导,花生四烯酸(arachidonic acid,AA)、二十二碳六烯酸(docosahexaenoic acid,DHA)和二十碳五烯酸(eicosapentaenoic acid,EPA)等长链多不饱和脂肪酸(10ng-chain polyunsaturated fatty acids,LC-PUFAs)的衍生物是一类重要的调控炎症的介质。炎性细胞间的交流和细胞内信号传递与LC-PUFAs有关。AA经环氧酶和脂氧合酶合成的类二十烷酸主要起促炎作用,但有的也有抗炎作用。DHA和EPA在体内起抗炎作用,由它们合成的消退素(resolvins,Rvs)和保护素(protectin,PD)是重要的抗炎活性物质。DHA和EPA还可以干扰炎性细胞内信号传导途径来抑制炎症反应。本文从炎症与疾病的关系、LC-PUFAs的衍生物及其促炎和抗炎机制等方面综述了AA、DHA和EPA在炎症中的作用。     

酸味酿造产品中三酸比例评析

酸味酿造产品中乳酸、醋酸、丁酸共存,但比例不同形成的酸味特征也不同。控制不同的环境条件,创造出不同的微生物区系,形成不同的三酸比例,才能形成不同的产品风格。该文对常见的酸味酿造产品中微生物区系的变化及三酸含量进行了分析。     

Growth of Aeromonas hydrophila K144 as Affected by Organic Acids

The influence of different acids on the aerobic growth kinetics of Aeromonas hydrophila was studied in BHI broth with 0.5 and 2.0% NaCl incubated at 5 and 19°C. Growth curve data were analyzed by the Gompertz equation and a nonlinear regression program; generation and lag times were calculated from the Gompertz parameters. Type of acid, pH, NaCl level and temperature influenced lag and generation times. The organic acids (acetic, lactic, citric and tartaric) inhibited growth at higher pH values than inorganic acids (HCl and H₂SO₄). The high NaCl level interacted with type of acid and pH to restrict growth of the organism at the lower temperature of incubation. Acetic and lactic acids were effective in controlling the growth of A. hydrophila and could readily be combined with low holding temperature to render foods free of the organism.     

杂多酸催化合成辛酸蔗糖酯

以蔗糖、辛酸为原料,杂多酸为催化剂合成辛酸蔗糖酯。用L16(45)正交设计优化实验,高效液相色谱法分析反应液组成。考察了催化剂种类和用量、反应温度、原料配比、反应时间等因素对辛酸蔗糖酯产率的影响,发现以二甲基亚砜为溶剂、蔗糖与辛酸摩尔比1∶9、磷钨酸用量为蔗糖质量的2.0%、110℃反应时间6h,蔗糖转化率达60%,产物产要是二酯。动力学研究发现,蔗糖反应级数为一级,反应表观速率常数为0.0059min-1(90℃)、0.0117min-1(110℃),反应表观活化能Ea=39.57kJ/mol。     

气相色谱法同时测定保健食品中的5种 不饱和脂肪酸

目的建立气相色谱法同时测定保健食品中亚油酸、α-亚麻酸、二十碳五烯酸(eicosapentaenoic acid,EPA)、二十二碳六烯酸(docosahexaenoic acid,DHA)和二十二碳五烯酸(docosapentaenoic acid,DPA)的含量。方法样品先采用氢氧化钾甲醇溶液进行皂化处理,再用三氟化硼甲醇溶液甲酯化,经HP-FFAP色谱柱(30m×0.53 mm,1.0μm)分离测定。结果 EPA甲酯、DHA甲酯、DPA甲酯、亚油酸甲酯、α-亚麻酸甲酯分别在0.03927~1.178、0.04200~1.260、0.03449~1.035、0.08368~1.255、0.08482~4.241 mg/mL的浓度范围内线性关系良好,相关系数r均大于0.999;检出限分别为0.0039、0.0042、0.0034、0.0042、0.0042 mg/mL;加标回收率在91.1%~109.3%之间,相对标准偏差均小于5%。结论该方法操作简单快捷,适用于保健食品中亚油酸、α-亚麻酸、EPA、DPA和DHA的测定。