酶膜反应器连续酶解花生蛋白的工艺研究

以花生蛋白为原料,Alcalase碱性蛋白酶为水解酶,研究了利用酶膜反应器连续酶解花生蛋白的最佳工艺条件.通过单因素实验选取实验因素与水平,并确定了操作压力为0.02MPa.再选取水解度(DH)为响应值,设计了四因素(pH、温度、底物浓度和加酶量)三水平的中心组合响应面实验.得出最佳工艺条件为:pH9.6,温度54℃,底物浓度2%,加酶量7440u/g.通过在最佳水解条件下进行水解,实际得到DH为26.13%.     

NADH氧化酶表达对干酪乳杆菌LC2W胞外多糖产量的影响

为了考察NADH氧化酶基因（nox）对于干酪乳杆菌LC2W胞外多糖（EPS）产量的影响,本研究以变异链球菌（Streptococcus mutans）基因组为模板,PCR扩增得到1374 bp的nox序列,并构建乳杆菌重组表达质粒p IB184-nox。通过电转化,构建得到一株过表达NADH氧化酶基因的重组干酪乳杆菌LC-nox。该重组菌在无氧发酵时,NADH氧化酶活力为0.7 U/m L,在有氧发酵时,NADH氧化酶活力达到2.65 U/mL。通过HPLC检测发现,发酵液中的乳酸含量随NADH氧化酶活力升高而降低。节省的碳源用于EPS合成,使得重组菌LC-nox在有氧条件下EPS产量最高达263.7 mg/L,比出发菌株提高75.4%。为乳杆菌EPS合成调控提供了新的理论基础。      

Production of gamma-aminobutyric acid from alcohol distillery lees by Lactobacillus brevis IFO-12005

Lactobacillus brevis IFO-12005 showed good growth in rice shochu distillery lees (kome shochu kasu). Almost all of the free glutamic acid (10.50 mM) in shochu kasu was converted to gamma-amino-butyric acid (GABA) within 2 d of stationary culture at 30 degrees C. The amount of GABA in the kome shochu kasu medium finally reached 10.18 mM. After centrifugation of the broth culture, the supernatant fraction was treated with a flocculation agent to form a clear solution, then passed through a column containing a synthetic adsorbents, SP-207 to remove the yellow pigment and flavors which are unnecessary from a sensory perspective. An economical and simple production process for GABA was established.     

Angiotensin I-converting enzyme inhibitor derived from soy protein hydrolysate and produced by using membrane reactor

Five different proteolytic enzymes, including Alcalase, Flavourzyme, trypsin, chymotrypsin and pepsin were employed to hydrolyze isolated soy protein (ISP) to produce the hydrolysates, respectively. The result indicated that hydrolysis of ISP for 0.5–6 h with Alcalase produced the highest ACE inhibitory activity. Therefore, Alcalase was selected for further study on optimization of hydrolysis conditions. The optimum conditions for Alcalase to hydrolyze ISP to produce the lowest IC₅₀ value were: E/S = 0.01, hydrolysis temperature = 50 °C, pH 9.0 and hydrolysis time = 6 h. Under these conditions, the IC₅₀ value of ISP was significantly reduced from 66.4 to 0.67 mg protein/ml. The lower IC₅₀ value represented the higher the ACE inhibitory activity. Moreover, several membranes with molecular weight cut-offs (MWCFs) of 1000–30,000Da were used to filter the hydrolysate. The 10 kDa permeate obtained from the treatment of the hydrolysate by 10,000 Da MWCF membrane could further reduce its IC₅₀ value from 0.668 to 0.078 mg protein/ml with a peptide recovery of 67.5%. An operation stability study showed that the membrane reactor system could maintain a steady production of ISP hydrolysate for over 8 h. The in vitro effect of gastrointestinal protease on ACE inhibitory activity of 10 kDa permeate was also investigated. The results suggested that gastrointestinal proteases have very little effect on the ACE inhibitory activity of 10 kDa permeate.     

Polymerization of proanthocyanidins catalyzed by polyphenol oxidase from lotus seedpod

This study aimed to investigate the profiles change in proanthocyanidins (PAs) catalyzed by polyphenol oxidase (PPO) from lotus seedpod in a model wine system (MWS). Results showed that PAs from lotus seedpod consisted of dimer (74.00 %) and trimer (22.75 %). PPO could tolerate ethanol concentrations below 20 % (v/v). The optimum temperature of PPO activity was 80 °C, and the optimal pH was 9.0. Its molecular weight was approximately 31 kDa, and its secondary structures were α-helix (59.0 %), β-sheet (4.3 %), turns (14.1 %), and random coils (22.6 %). In the MWS, the trimers gradually increased from 22.55 % at 0 h (control) to 100 % at 10 h incubation, while the dimers decreased from 74.33 % (control) to 0 % at 10 h incubation. Moreover, the composition of the precipitate formed at different incubation time points was approximately 16.54 % of monomers, 21.03 % of dimers, and 62.43 % of trimers at 2 h incubation with PPO. The results from this study have provided in vitro evidence for a possible application of PPO in red wine aging.     

Preparation of elemental selenium-enriched fermented milk by newly isolated Lactobacillus brevis from kefir grains

A Lactobacillus brevis strain CGMCC No. 6683 that can survive at high selenium concentrations was isolated from kefir grains. Using a scanning electron microscope equipped with an energy dispersive X-ray spectroscope, it was shown that L. brevis CGMCC No. 6683 reduced sodium selenite into elemental selenium. L. brevis CGMCC No. 6683 was then pre-treated with sodium selenite to prepare selenium-enriched L. brevis. To study the selenium-enriched yoghurt, selenium-enriched L. brevis was co-fermented in skimmed milk with traditional yoghurt starter culture (Streptococcus thermophilus and Lactobacillus bulgaricus). Results showed that it enhanced selenium concentration in the selenium-enriched yoghurt, and more importantly, elemental selenium was detected in the selenium-enriched yoghurt. The isolated L. brevis provides a new way of preparing a functional dairy product: elemental selenium-enriched fermented milk.     

酶膜反应器在蛋白酶解过程中的研究进展

介绍酶膜反应器的基本概念、原理、特点和分类。重点阐述酶膜反应器作为一种新型的酶解反应装置在水解蛋白过程中的应用及其突出的优势,同时也提出了该装置应用的局限性。     

Enhanced production of compound K in fermented ginseng extracts by Lactobacillus brevis

Food Science and Biotechnology - The purpose of this study is to establish the best condition and microorganism for preparation of fermented ginseng including rich compound K. When raw ginseng...     

Production of angiotensin-converting enzyme inhibitory peptides in Iranian ultrafiltered white cheese prepared with Lactobacillus brevis KX572382

In this study, ultrafiltered (UF) Iranian white cheese made with adjunct cultures including six Lactobacillus isolates (Lactobacillus brevis, L. casei and L. plantarum) from traditional Iranian Motal cheese. The peptide extract (<5 kDa) of cheese samples were assessed for angiotensin-converting enzyme (ACE)-inhibitory activity during ripening (5 °C). Among the strains used, L. brevis KX572382 (M8) was selected because of the greater increase in (ACE)-inhibitory activity in the cheese (P < 0.05). The highest activity of M8 extract was observed on the 28th (71.72%) day of ripening (P < 0.05). Proteolytic activity assessment and RP-HPLC peptide profile of M8 water-soluble extracts (WSEs) indicated the effect of M8 on further protein degradation due to secondary proteolysis. A total of 7 different peptide sequences, previously known in the literature for their ACE-inhibitory activity, were tentatively identified by LC/ESI-MS in 28-day M8 peptide extract. Although the effect of M8 on pH and the proteolysis development in cheese was significant, no adverse effect was observed on the sensory properties. In conclusion, M8 strain can enhance the functional properties of Iranian UF white cheese.     

生物酶催化废白土油制备生物柴油的工艺研究

研究生物酶催化废白土油与甲醇酯交换制备生物柴油的最佳工艺条件。通过对比相当用量的Lipozyme TL IM和Novozyme 435的催化效果,筛选出Lipozyme TL IM为适宜的酶;在此基础上,以醇油摩尔比、生物酶添加量、反应温度、反应时间为自变量,生物柴油得率为响应值,进行酯交换制备生物柴油的响应面优化实验。结果表明,酯交换反应最佳条件为：醇油摩尔比4∶1,Lipozyme TL IM添加量10%(以废白土油质量计),反应温度35℃,反应时间15 h;在此条件下,生物柴油得率为95.9%,所得生物柴油非常接近0#柴油的质量标准。     

Determination of total cholesterol content in food by flow injection analysis with immobilized cholesterol oxidase enzyme reactor

A new analytical method has been developed--including a new sample preparation procedure--for the automatic determination of total (free and bound) cholesterol in food by flow injection analysis (FIA) with immobilized (cholesterol oxidase) enzyme reactor (IMMER). A suitable sample preparation procedure has been applied to eliminate the problems derived from sensitivity of FIA equipment to common organic solvents used for dissolving of cholesterol: after direct saponification the non-saponificable fraction was dissolved in water phase detergent (sodium cholate) solution. The analytical method is based on the oxidation by cholesterol oxidase followed by the photometric determination of hydrogen peroxide (at 500 nm) using the indicator reaction with peroxidase. The new FIA method was tested for commercial food samples such as whole egg powder and dried pasta. The results were compared with data obtained by GC-determination. It was found that this new procedure is suitable for rapid automated measurement of total cholesterol content in foodstuff and consequently, the FIA technique with immobilized enzyme reactor could be an alternative to the widely used gas-chromatographic (GC) method.     

短乳杆菌鸟氨酸氨甲酰基转移酶对黄酒发酵中氨基甲酸乙酯的调控作用

氨基甲酸乙酯是存在于黄酒发酵过程中的一种潜在致癌物质.本文研究添加鸟氨酸氨甲酰基转移酶对氨基甲酸乙酯及黄酒基础品质的影响.根据鸟氨酸氨甲酰基转移酶可以催化、降解氨基甲酸乙酯的前体物瓜氨酸的原理,采用基因工程手段从短乳杆菌中克隆并大量表达鸟氨酸氨甲酰基转移酶,经Ni-NTA琼脂糖纯化树脂纯化后添加到黄酒发酵的不同阶段.结...     

Continuous production of L-carnitine with NADH regeneration by a nanofiltration membrane reactor with coimmobilized L-carnitine dehydrogenase and glucose dehydrogenase

L-carnitine dehydrogenase (CDH) was partially purified from Pseudomonas putida IAM12014 for the stereospecific reduction of 3-dehydrocarnitine to L-carnitine. CDH and glucose dehydrogenase (GDH) were coimmobilized in a nanofiltration membrane bioreactor (NFMBR) for the continuous production of L-carnitine from 3-dehydrocarnitine with NADH regeneration. In the NFMBR, NAD was partially immobilized through rejection by the nanofiltration membrane and effectively regenerated by the conjugation reaction of CDH and GDH. Since 3-dehydrocarnitine was unstable at neutral pH, it was maintained under acidic conditions (pH 0.7) and supplied to the NFMBR separately from the other substrates, glucose and coenzyme NAD. As 50 mM 3-dehydrocarnitine in HCl solution, 0.05 mM NAD, and 100 mM glucose in 0.5 M Tris buffer (pH 8) were continuously supplied to the NFMBR with immobilized CDH (200 U/ml) and GDH (200 U/ml) at the retention time of 80 min and temperature of 25 degrees C, the maximum conversion, reactor productivity, and NAD regeneration number were 78%, 113 g/l/d, and 780, respectively. The half-life of the NFMBR was longer than 500 h.     

一株从泡菜中分离的产细菌素乳杆菌的鉴定及细菌素特性研究

从泡菜中分离的329 株乳酸菌中,筛选出一株产细菌素的乳酸菌,编号为SD-22。经生理生化实验鉴定为短乳杆菌(Lactobacillus brevis)。在排除过氧化氢干扰和有机酸抑菌作用后,brevicin SD-22 对革兰氏阳性细菌和革兰氏阴性细菌具有较好的抑制作用,对部分真菌也有抑制作用,但对酵母菌和青霉无抑制作用,具有良好的热稳定性和pH 值稳定性,经胰蛋白酶、胃蛋白酶和蛋白酶K 处理后抑菌活性完全消失,对α- 淀粉酶不敏感。因此初步认为该菌株是一株产广谱细菌素的乳酸菌。     

一株曲拉源短乳杆菌的分离及其体外抑菌特性分析

摘要：目的 天然牦牛乳曲拉中蕴含着大量的乳酸菌，为丰富食品发酵的有益菌种资源，本试验分离筛选具有抑菌活性的乳酸菌。方法 以西藏牦牛乳曲拉为分离基质，MRS(De Man Rogosa Sharpe)加上1%CaCO3为选择鉴别培养基，运用形态学观察、生理生化试验、16S r RNA基因序列、药敏试验对分离菌株进行菌种鉴定，通过抑菌试验探究其抑菌活性。结果 分离菌株NWMCC0322被鉴定为短乳杆菌，其对青霉素类、四环素类、大环内酯类、酰胺醇类、喹诺酮类、磺胺类、头孢类药物敏感，对多烯类药物中敏。NWMCC0322对指示菌金黄色葡萄球菌的抑菌效果最好，其抑菌圈直径达12.0±0.6 mm，其次为甲型副伤寒沙门氏菌11.3±0.3 mm，大肠杆菌11.0±0.5 mm。结论 该短乳杆菌对食品污染常见菌株具有抑菌效果，在食品发酵中有较好的应用价值。     

Purification of goat beta-lactoglobulin from whey by an ultrafiltration membrane enzymic reactor

This paper presents a novel contribution to the purification of goat beta-lactoglobulin by using an ultrafiltration membrane enzymic reactor. The basis of the purification process was the enzymic hydrolysis of contaminating proteins, alpha-lactalbumin and traces of serum albumin, by pepsin at 40 degrees C and pH 2, conditions under which beta-lactoglobulin is resistant to peptic digestion. Simultaneously, beta-lactoglobulin and peptides were separated by ultrafiltration. beta-Lactoglobulin was retained in the reactor while peptides generated by hydrolysis from alpha-lactalbumin and serum albumin permeated through the membrane. The process was made continuous by the addition of fresh whey to replace the lost permeate. Three mineral membranes with 10, 30 and 50 kDa molecular mass cut-off were tested and the 30 kDa membrane was selected for the continuous process. The simultaneous purification and concentration of beta-lactoglobulin from clarified goats' whey was achieved in a single step. The ultrafiltration membrane enzymic reactor could treat eight reactor volumes of clarified whey. The recovery of beta-lactoglobulin was 74%, its purity was 84% and its concentration 6.6-fold that in the initial clarified whey.     

乳源短乳杆菌M8 S- 层蛋白的提纯及其生物学特征分析

通过利用原子力显微镜(AFM)来观察凝胶过滤层析法提纯短乳杆菌M8菌株的S-层蛋白的表面形貌,同时探讨S-层蛋白的再生特性及黏附特性。结果表明：凝胶过滤层析法能够获得纯度较高的S-层蛋白;该蛋白在纯水中可自我组装成纳米级“团簇”结构;去除S-层蛋白的菌体细胞仍然具有生命活性,适当培养后可重新表达该蛋白;短乳杆菌M8可黏附到Caco-2细胞上,其S-层蛋白介导此过程。