生物胺降解乳酸菌的筛选与特性研究

为筛选出高效降解生物胺的优良乳酸菌,该研究以实验室前期筛选的41株乳酸菌作为研究对象,运用显色培养、高效液相色谱（HPLC）法对菌株生物胺产生和降解能力进行筛选,并对筛选出的菌株进行耐盐及耐酸实验。结果表明,筛选得到5株高效降解生物胺菌株,其中以植物乳杆菌（Lactobacillus plantarum）30的生物胺降解能力最好,其对尸胺、组胺、酪胺的降解率分别为62.42%、74.32%、89.97%。植物乳杆菌30能在含盐量0～9%和pH为4.5～8.5环境中较好的生长繁殖,该菌株无生物胺生成活性,并同时具备生物胺降解能力和耐盐耐酸能力,可作为蛋白质类发酵食品的发酵剂运用于食品中降低生物胺毒性。     

不同条件对两株乳酸菌降解生物胺的影响

为探究从新疆熏马肠中分离出的植物乳杆菌和唾液乳杆菌在不同环境条件下降解生物胺的效果,通过高效液相色谱(High Performance Liquid Chromatography,HPLC)检测两株菌在不同条件下(生物胺浓度、初始pH、温度、NaCl浓度和乙醇浓度)降解生物胺的情况。结果表明:当生物胺浓度在200~300 μg/mL时两株菌生物胺降解能力基本达到最大,而当生物胺浓度大于300 μg/mL时两株菌降解生物胺的能力逐渐下降,但唾液乳杆菌(Lactobacillus salivarius)对生物胺的降解能力显著高于植物乳杆菌(Lactobacillus plantarum)(p<0.05),分别在pH为6.5和7.0时对大多数生物胺具有较高的降解能力;两株菌均在32~37 ℃温度范围内对生物胺有较好的降解效果;当NaCl浓度达到8%,乙醇浓度达到4.5%时,两株菌对生物胺的降解活性被强烈抑制。可见本研究中的唾液乳杆菌相对于植物乳杆菌更适合作为商业发酵剂,具有更好的降解生物胺的特性。     

葡萄酒酿造中降解生物胺乳酸菌的筛选、鉴定及其特性

葡萄酒发酵过程中乳酸菌脱羧形成生物胺,当生物胺摄入过量时会对人体造成危害。为筛选具有降解生物胺能力的乳酸菌,采用生物胺显色培养基和高效液相色谱法筛选获得1株降解生物胺能力较好的菌株,经形态学和分子生物学鉴定为植物乳植物杆菌,并命名为NXU-Q12。在此基础上,进一步研究其生长特性和降解生物胺的能力,其在不同pH和乙醇浓度下均可生长,但在pH为3～4.5,乙醇体积分数为10%～14%时生长活性和降解生物胺的能力同时受到抑制。文章为葡萄酒发酵过程中生物胺的调控提供优良菌株和一定的参考价值。     

泡菜中乳酸菌的分离鉴定及抗性筛选

该研究从泡菜中分离乳酸菌,通过形态观察及分子生物学技术对其进行鉴定,并对其进行人工胃液和胆盐耐受性试验,以期筛选性能优良乳酸菌。结果表明,从泡菜中共分离出71株乳酸菌,经鉴定分别为消化乳杆菌（Lactobacillus alimentarius）32株、植物乳杆菌（Lactobacillus plantarum）27株、发酵乳杆菌（Lactobacillus fermentium）1株、棒状乳杆菌（Lactobacillus coryniformis）2株、有害片球菌（Pediococcus damnosus）9株。其中,植物乳杆菌和发酵乳杆菌可用于食品,且植物乳杆菌S74和S78具有较高的抗人工胃液能力,存活率分别为（94.73±4.56）%、（108.73±7.16）%;菌株S74在0.3%的胆盐中的生长效率[（7.41±3.28）%]低于菌株S78[（10.04±4.90）%]。说明植物乳杆菌S78在pH 3.0的人工胃液和0.3%的胆盐环境均具有良好的耐受能力,在功能性泡菜及益生菌产品方面具有一定的开发潜力。     

萝卜泡菜母水中乳酸菌分离鉴定与发酵特性比较

从四川传统萝卜泡菜母水中分离出优势乳酸菌,对其进行鉴定和发酵特性研究。用纯培养的方法对菌株进行分离纯化,经生理生化实验和16S rDNA测序对菌种进行鉴定。对照类肠膜魏斯氏菌(Weissella paramesenteroides),研究其生长特性、耐受性和降解亚硝酸盐能力,评价筛选菌株的发酵特性,并采用气相色谱-离子迁移谱对菌株产挥发性风味物质的能力进行检测分析。筛选出L2、L3、L7、L15和L24等5株菌分别为：布氏乳杆菌(Lactobacillus buchneri)、短乳杆菌(Lactobacillus brevis)、乳酸片球菌(Pediococcus acidilactici)、植物乳植杆菌(Lactiplantibacillus plantarun)和鼠李糖乳酪杆菌(Lacticaseibacillus rhamnosus)。L15和L24生长速度比其他菌株快;菌株L7、L15和L24产酸能力强;菌株L15和L24对NaCl的耐受性最好;菌株L7的NaNO₂耐受性最差;5种菌株降解亚硝酸盐的能力均高于90%。5株乳酸菌产挥发性物质的特征不同,L3产...     

用于低脂干酪开发的附属发酵剂的筛选

目的筛选出生产低脂干酪所需添加较为适宜的附属发酵剂。方法选择7株乳酸菌作为研究对象,测定其总肽酶活力、菌株生产能力、产酸能力、产粘能力、菌体自溶度、产胞外多糖数量、蛋白质水解能力等7种指标。结果 7株乳酸菌中嗜酸乳杆菌(Lactobacillus acidophilus,LA)总肽酶活力最高,为(256.46±11.88)μg/L,植物乳杆菌(Lactobacillus plantarum,LP)最低;菌株生长能力LA最好;菌株副干酪乳杆菌(Lactobacillus paracasei,LPC)在发酵10 h后的产酸能力最强且趋于稳定,pH在3.8左右;LP菌株产粘能力最优,为(168.10±14.72)mPa·s;瑞士乳杆菌(Lactobacillus helveticus,LH)菌株自溶度最好,达到35%;产胞外多糖数量最优菌株是LP,含量为(792.69±35.94)mg/L;LH的蛋白质水解能力最高,水解产生的游离氨基酸含量为(94.78±2.82)mg/L。结论 LH作为低脂干酪的附属发酵剂较好。     

高效降解生物胺乳酸菌的筛选、鉴定及特性研究

为获得高效降解生物胺的乳酸菌，以鱼露为试验原料，从鱼露中分离纯化并筛选具有高效降解4种常见生物胺活性的菌株，通过形态观察、碳源利用和分子生物学鉴定，该菌株，并研究其生长和生物胺降解特性。结果表明，共筛选得到8株生物胺降解菌株，其中最佳生物胺降解菌为FSCBAD033，该菌株被鉴定为发酵柠檬乳杆菌（Limosilactobacillus fermentum）。该菌株可以降解86.4%腐胺、78.5%尸胺、72.3%组胺和100%酪胺，且在含有前体氨基酸的培养基中不积累该四种生物胺。菌株FSCBAD033的最适生长温度、初始pH值和NaCl含量分别为40 ℃、6和3%，其在温度30～40 ℃、初始pH值5～7、NaCl含量不超过6%的范围内降解生物胺能力较高（四种生物胺降解率均＞50%）。     

传统发酵牦牛酸乳中益生性乳酸菌的体外筛选

为获得具有潜在益生特性的乳酸菌,实验以抗人工胃液、胆盐中生长效率和细胞表面疏水性为筛选指标,对羊八井地区传统发酵牦牛酸乳中分离出的38株乳酸菌进行体外筛选。最终通过测定结果综合比较得出菌株编号为YBJ-3、YBJ-11、YBJ-15、YBJ-17、YBJ-30和YBJ-33乳酸菌的益生特性较好,其中4株乳酸菌抗人工胃液能力大于90%,在胆盐中良好生长。通过16S r DNA序列分析鉴定,菌株编号YBJ-3为屎肠球菌(Enterococcus faecium)、YBJ-11为发酵乳杆菌(Lactobacillus fermentum)、YBJ-15为耐久肠球菌(Enterococcus durans)、YBJ-17为植物乳杆菌(Lactobacillus plantarum)、YBJ-30为植物乳杆菌(Lactobacillus plantarum)、YBJ-33为干酪乳杆菌(Lactobacillus casei)。     

泡菜中亚硝酸盐降解乳酸菌的筛选与鉴定

目的从泡菜中筛选亚硝酸盐降解能力强的乳酸菌。方法采用平板涂布法和平板划线法从泡菜中分离若干株产酸菌株,从中筛选出产酸能力强、生长速度快、亚硝酸盐降解性能高的菌株。对优选菌株进行了形态学、生理学鉴定,测定其16S rRNA基因序列,并检测其耐盐能力、糖谱、产酸能力、抑菌性能及亚硝酸盐降解能力。结果该产酸菌株为植物乳杆菌(Lactobacillus plantarum),此菌在37℃、7.0%NaCl及0.1%NaNO2条件下生长良好;37℃MRS培养基培养24 h后pH下降为3.96;该菌株在以葡萄糖、蔗糖、果糖和菊粉为碳源的LB培养基中生长良好;具有较高的乳酸生产能力,对亚硝酸盐的降解率高达91.3%。结论从泡菜中筛选的乳酸菌株具有较高的耐盐能力及亚硝酸盐降解能力。     

西北地区泡菜中乳酸杆菌的生物学特性

从西北地区采集的泡菜中分离出四株乳酸菌分别是植物乳杆菌(Lactobacillus plantarum)、短乳杆菌(Lactobacillus brevis)、副干酪乳杆菌(Lactobacillus paracasei)和戊糖乳杆菌(Lactobacillus pentose)。37℃MRS液体培养基中静置培养一定时间,分别测定了四种乳酸杆菌的产酸能力、硝酸盐还原性、亚硝酸盐的降解能力、耐酸耐盐能力和生长特性。结果表明:LPL,LPA和LPE培养15h到达稳定期,LBR培养27h到达稳定期;四株乳酸杆菌的总酸含量分别为1.98%,1.97%,2.10%和1.77%,亚硝酸盐降解率依次为0.87%,0.88%,0.93%和0.88%;四株菌的硝酸盐还原性都为阴性,最低耐酸性为pH 5,最高耐盐性为NaCl含量4%。     

鱼露中组胺降解菌的分离筛选及耐受性

生物胺(biogenic amines)在某些食品尤其是发酵食品中广泛存在,具有一定的食用安全隐患.为获得用于鱼露等发酵食品的生物胺降解菌,从天然发酵鱼露中采用双层显色培养基法初步筛选出不产生物胺的菌株,再用高效液相色谱(HPLC)法进行复筛,得到一株具有高效组胺降解能力的菌株MZ5.经鉴定该菌株为库德毕赤酵母(Pic...     

Screening selected strains of probiotic lactic acid bacteria for their ability to produce biogenic amines (histamine and tyramine)

The aim of this study was to investigate the production of biogenic amines (BA), histamine and tyramine by some probiotic lactic acid bacteria (LAB). Fifteen strains representing six LAB species were screened qualitatively by growing them in a decarboxylase medium. Quantitative analysis was carried out by HPLC analysis with direct derivatization of acid extracts. Lactobacillus casei (TISTR 389) and Lactobacillus delbrueckii subsp. bulgaricus (TISTR 895) were found to produce BA. The highest levels of histamine (1820.9 ± 3.5 mg L^?1) and tyramine (5486.99 ± 47.6 mg L^?1) formation were observed for the TISTR 389 strain, while TISTR 895 produced only histamine (459.1 ± 0.63 mg L^?1) in the decarboxylase broth. Biogenic amine potential was not observed for the Lactobacillus acidophilus, Lactobacillus lactis subsp. lactis, Lactococcus lactis subsp. lactis, and Lactobacillus plantarum strains studied. This study confirmed that BA formation is strain dependent and not related to the species. Therefore, careful screening for amino acid decarboxylase activity is recommended before selecting LAB as appropriate starter or probiotic strains in food and dairy industry.     

浓香型大曲中降解生物胺菌株的筛选及应用

从浓香型白酒大曲中筛选出生物胺降解菌,并研究其生长特性和在固态发酵条件下降解生物胺的能力。利用高效液相色谱法测定发酵液中生物胺含量,得到1株降解生物胺效果最好的酵母菌,经形态学和分子生物学鉴定其为奥默柯达酵母,将其命名为Kodamaea ohmeri HJM。研究K.ohmer HJM的生长特性,得到其最高耐受温度为43℃,乙醇最高耐受体积分数为14%,耐受酸最低为pH 2,并具有较高的葡萄糖耐受性。在固态发酵条件下,利用小麦为发酵基质得到接种菌液样品组和空白对照组中生物胺含量具有显著差异(P<0.05),其对几种生物胺的降解率分别为:腐胺(39.19±0.08)%、尸胺(33.77±0.06)%、甲胺(32.44±0.06)%、乙胺(23.39±0.06)%、吡咯烷(63.42±0.02)%、异戊胺(49.83±0.07)%、环己胺(49.73±0.03)%、环戊胺(66.07±0.08)%。该研究从浓香型大曲中分离出对生物胺降解率较高的菌株,可为白酒中生物胺的调控提供参考价值。     

Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

Isolation,characterization, and application of biogenic amines-degrading strains from fermented food

The aim of the present study was to screen a type of microorganism for degradation of bioamines in fermented food, investigate their characteristics in growth and degrading bioamines, and also assess the effects of inoculating strain starter cultures on biogenic amines (BAs) accumulation in soy sauces. The screening for no-BAs-producing strains was carried out using a double color method and an oxidase test. The degradation rates were determined by high-performance liquid chromatography with fluorescence detection (HPLC-FLD). In this study, strains (J2 and J3) with high ability were isolated from some naturally fermented food samples. They were identified as Wickerhamomyces anomalus and Millerozyma farinosa, respectively, by partial amplification of the 16S rRNA gene. The strains exhibited high multiple BAs degradation activity over a range of pH values (5–8), temperatures (28–35°C), salt (NaCl) concentrations (0–10%), and ethanol concentrations (0–10%). One of the isolated strains, W. anomalus, has the highest biodegradability for putrescine, cadaverine, histamine, and tyramine with degradation rates of 61, 66, 67, and 59% in an experimental model, and 39.3, 33.6, 39.9, and 43.1% in fermented soy sauce containing 10% of NaCl, respectively. Additionally, the study of mechanism demonstrated that the removal ability of BAs was mainly due to biodegradation.     

Selection of a versatile Lactobacillus plantarum for wine production and identification and preliminary characterisation of a novel histamine-degrading enzyme

In this study, a desirable and versatile Lactobacillus plantarum strain possessing great ability of L-malic acid consumption, biogenic amine degradation and resistance to wine harsh environment was obtained through successive screenings. Malolactic fermentation (MLF) in contaminated grape wine and cherry wine (supplemented with histamine, tyramine and cadaverine) conducted by this strain was finished within 24 and 18 days, respectively, and the concentration of histamine, tyramine and cadaverine was decreased by over 57% following MLF. The enzyme from L. plantarum responsible for the amine degradation was purified to homogeneity by four steps including cell disruption, ammonium sulphate fractionation, an anion chromatography and a gel filtration chromatography. Such enzyme was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) after N-terminal sequence analysis and protein Blast, a tetramer enzyme with a subunit molecular weight of 36 kDa. The optimum values of pH and temperature of this enzyme were at pH 7.5 and 40 °C, and stable between pH 5.5–8.5 and 30–50 °C.     

黄酒中不产生物胺乳酸菌的筛选及应用

为筛选不产生物胺的乳酸菌菌株,对黄酒发酵醪液及米浆水中的微生物进行培养及与分离纯化,发现3株菌不具有氨基酸脱羧酶活性,但具有一定的生物胺降解能力。使用黄酒发酵液培养该3株菌,发现在2.50%Vol的黄酒培养液中,3株菌能正常生长;在8.80%Vol黄酒培养液中,菌株5-4和8-3生长能力明显弱于14-2-1;在15.70%Vol黄酒培养液中,3株菌均受到严重抑制。菌株14-2-1生长速率最快,繁殖最旺,经鉴定为植物乳杆菌,同时菌株产酸能力较强,产酸速率也较快,表现出一定的优越性,应用于黄酒酿造中,能有效降低黄酒中生物胺的含量。