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%）。     

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...     

豆瓣酱生物胺降解菌株的筛选、鉴定及其降解特性研究

为保障传统发酵食品的质量安全,从豆瓣酱样品中筛选生物胺降解菌株,并研究其在不同盐浓度、温度、pH下对组胺和酪胺(毒性最强的2种生物胺)的降解特性,以便于更好地应用于传统发酵食品。通过研究菌株在磷酸盐缓冲液和在发酵液中对生物胺的降解率以及在发酵液中产生生物胺的能力,最终筛选得到1株具有较强生物胺降解能力的乳酸片球菌(Pediococcus acidilactici)M-28。对该菌株在不同条件下降解组胺和酪胺的能力进行分析,发现P.acidilactici M-28在盐质量浓度为0～90 g/L时对组胺和酪胺均具有较高的降解率,最高降解率分别达到89.02%和31.49%;该菌株降解生物胺的最适温度为30℃,在该条件下对组胺和酪胺降解率分别为65.51%和28.06%;最适pH值为6,在该条件下对组胺和酪胺的降解率分别为62.73%和23.78%。     

复合菌株发酵对鱼露中生物胺含量的影响

文中研究了以米曲霉为种曲发酵鱼露的工艺,在此基础上,接人木糖葡萄球菌、植物乳杆菌及其复合菌株共同发酵,分析了单一菌株和复合菌株对鱼露中生物胺(组胺、腐胺、酪胺、亚精胺)生成量的影响.结果表明,单一的木糖葡萄球菌及其与植物乳杆菌的复合菌株都可以降低鱼露中生物胺的含量,其中,在米曲霉生长的稳定期时接入复合菌株时效果最好,与对照组相比总胺降低了61.9%.     

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。     

Limosilactobacillus fermentum FSCBAD033控制带鱼加工下脚料发酵鱼露中生物胺含量研究

目的 控制带鱼加工下脚料低盐发酵鱼露中的生物胺含量。方法 以耐盐性高效生物胺降解乳酸菌Limosilactobacillus fermentum FSCBAD033为功能发酵剂控制带鱼加工下脚料发酵鱼露中的生物胺含量,并研究其对鱼露发酵过程中的pH值、NaCl含量、总可溶性氮(Total Soluble Nitrogen,TSN)含量、氨基酸态氮(Amino Acid Nitrogen,AAN)含量、挥发性盐基氮(Total Volatile Basic Nitrogen,TVB-N)含量以及发酵结束时感官风味的影响。结果 与Control组相比,接种L. fermentum FSCBAD033发酵剂对鱼露发酵过程中的NaCl含量影响不大(p>0.05),但可促进pH值下降以及TSN和AAN含量上升,并显著抑制TVB-N的产生(p<0.05)；显著降低发酵结束时鱼露中52.38%尸胺、40.21%组胺、45.44%酪胺、23.74%腐胺、52.67%苯乙胺和43.26%总生物胺(p<0.05),对含量较低的色胺、精胺和亚精胺影响不大；此外,接种L. fermentum FSCBAD033发酵剂可增强发酵结束时鱼露的鲜味和肉味,减弱氨味、腥味和臭味,整体提升鱼露感官风味。结论 L. fermentum FSCBAD033可作为带鱼加工下脚料低盐发酵的功能发酵剂,用于生产低盐、营养丰富、感官风味好且生物胺含量低的高品质鱼露产品。     

温度和姜精油对金枪鱼品质影响及生物胺相关性

将分别贮藏于不同温度(0、4、15℃)条件下的金枪鱼样品,以及4℃条件下用姜精油作为生物抗氧化剂处理过的样品作为研究对象,定期测定金枪鱼生物胺(腐胺Put、尸胺Cad、组胺His、酪胺Tyr)、反式尿刊酸的含量、挥发性盐基氮(total volatile basic nitrogen,TVB-N)值、K值和菌落总数,研究温度和姜精油对金枪鱼生物胺含量的影响及组胺与反式尿刊酸的关系。结果表明,温度对金枪鱼生物胺生成有较大影响,0℃条件下生物胺含量显著低于其他贮藏温度,到第6天时组胺、酪胺、腐胺和尸胺含量分别为28.25、5.47、14.84 mg/kg和17.05 mg/kg,而此时15℃条件下金枪鱼组胺、酪胺、腐胺和尸胺含量分别为135.4、14.63、29.49 mg/kg和41.55 mg/kg;生物胺中组胺含量最高,而酪胺含量始终处于低水平状态,即使在15℃条件下到第7天时其含量仍为16.24 mg/kg;结果同时表明,生物抗氧化剂对生物胺的产生和微生物生长有一定的抑制作用,并能有效地延缓蛋白质和ATP降解。相关分析和回归分析表明,组胺、酪胺、尸胺、与TVB-N值、K值和菌落总数高度相关,其相关系数r在0.804~0.981之间,且生物胺和菌落总数、TVB-N值之间存在重要对应关系,反式尿刊酸对组胺的产生影响不大,两者之间相关性较弱,在4℃时其相关系数r为0.630。     

一株具有降生物胺能力葡萄球菌的筛选及其降解特性

近年来，发酵肉制品因其营养丰富、风味独特等优势而在市场中具备一定优势，但发酵肉中常存在生物胺含量超标等卫生问题，易引发了人们对其安全性的恐慌。该研究通过双层显色培养基初筛、高效液相色谱复筛的方法，再结合菌株的生物胺氧化酶活力大小与生长特性，从传统发酵肉中筛选出一株针对8种常见的生物胺均有一定降解作用的菌株aM，经16S rDNA与糖酵解试验鉴定为沃氏葡萄球菌（Staphylococcus warneri）。该菌株在Luria-Bertani（LB）液体培养基中37 ℃培养24 h后，对生物胺总降解率为18.78%，对苯乙胺、色胺及酪胺的降解率分别为28.56%、23.16%和22.45%。此外，还探究了aM不同环境条件下生物胺降解率的影响，发现其在温度30~40 ℃，pH值7及盐质量浓度为0%~6%时降解率最高，这为之后菌株的实际生产应用中优化发酵条件提供了思路。     

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。     

甜瓣子中生物胺产生菌株和降解菌株的筛选

甜瓣子发酵是豆瓣酱生产的重要阶段,低盐发酵甜瓣子中生物胺含量极易超标。采用生物胺显色培养基,以生物胺为唯一氮源的培养基(BAs)进行筛选,采用高效液相色谱法定量分析和荧光检测定性分析方法测定生物胺含量,并通过16S rDNA、18S rDNA和ITS测序鉴定菌株。结果表明,甜瓣子中产胺菌株多为芽孢杆菌属、肠杆菌属、短杆菌属,少数小孢根霉等也产生物胺,其中枯草芽孢杆菌B11在9%和12%盐度下发酵16 d,分别产苯乙胺233.9,546.5 mg/kg。降胺菌株以美极梅奇酵母属、片球菌属、乳杆菌属为主,其中香肠乳杆菌F003在6%盐度发酵的甜瓣子中发酵16 d,可完全降解100 mg/L的苯乙胺,对100 mg/L的组胺、酪胺和精胺的降解率分别达57.61%,25.52%和45.99%。F003回接至6%的甜瓣子,发酵得到的甜瓣子酯香味浓,无刺激性气味,甜瓣子品质好。在发酵甜瓣子体系中,枯草芽孢杆菌是产生物胺的主要菌种。本研究筛选到1株高效降解生物胺的香肠乳杆菌F003,可为甜瓣子发酵体系中生物胺的控制提供菌株支持。     

Inhibitory Effects of Spices on Biogenic Amine Accumulation during Fish Sauce Fermentation

The presence of high levels of biogenic amines is detrimental to the quality and safety of fish sauce. This study investigated the effects of ethanol extracts of spices, including garlic, ginger, cinnamon, and star anise extracts, in reducing the accumulation of biogenic amines during fish sauce fermentation. The concentrations of biogenic amines, which include histamine, putrescine, tyramine, and spermidine, all increased during fish sauce fermentation. When compared with the samples without spices, the garlic and star anise extracts significantly reduced these increases. The greatest inhibitory effect was observed for the garlic ethanolic extracts. When compared with controls, the histamine, putrescine, tyramine, and spermidine contents and the overall biogenic amine levels of the garlic extract‐treated samples were reduced by 30.49%, 17.65%, 26.03%, 37.20%, and 27.17%, respectively. The garlic, cinnamon, and star anise extracts showed significant inhibitory effects on aerobic bacteria counts. Furthermore, the garlic and star anise extracts showed antimicrobial activity against amine producers. These findings may be helpful for enhancing the safety of fish sauce.     

Biogenic amine production by Oenococcus oeni during malolactic fermentation of wines obtained using different strains of Saccharomyces cerevisiae

Twenty-six wild Oenococcus oeni strains were investigated for their ability to form biogenic amines during malolactic fermentation in synthetic medium and in wine. Eight strains produced histamine and tyramine in screening broth at concentrations of 2.6-5.6 mg/L and 1.2-5.3 mg/L, respectively. Based on their ability to form biogenic amines, five strains were selected to inoculate three wines obtained by the fermentation of three different Saccharomyces cerevisiae strains (A, B, and C). All bacterial strains could perform malolactic fermentation for short periods in wine C, whereas only one strain performed complete malolactic fermentation in wines A and B. Two O. oeni strains (261 and 351) produced histamine and tyramine in wine C. Time-course analysis of these compounds showed that for both strains, histamine and tyramine production began at day 10 and finished on day 25, after the end of malolactic fermentation. These results indicate that the ability of O. oeni to produce histamine and tyramine is dependent on the bacterial strain and on the wine composition, which in turn depends on the yeast strain used for fermentation, and on the length of bacteria-yeast contact time after the completion of malolactic fermentation.     

Novel starter cultures to inhibit biogenic amines accumulation during fish sauce fermentation

Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.     

不同贮藏温度下鲐鱼生物胺变化的研究

为探究不同温度和预处理对鲐鱼生物胺含量变化的影响,用高效液相色谱法分析了整鱼和去内脏鱼在不同贮藏温度下8种生物胺以及总生物胺含量的变化,结果发现,鲐鱼中主要的生物胺为组胺、腐胺、尸胺、酪胺。随着温度的升高,鲐鱼中总生物胺含量的增加迅速变快。在0、4、10、15、20、25和30 ℃贮藏1 d后,整鱼中总生物胺的积累量分别为40.34、93.44、107.95、73.39、119.99、4649.90、6446.43 mg/kg。在0、4、10、15、20、25和30 ℃贮藏条件下,整鱼中组胺的含量超过国家规定的高组胺鱼类安全限量(400 mg/kg)的时间分别为12、5、4 d和48、36、15、14~16 h,-18 ℃贮藏6个月,组胺积累量仅为13.45 mg/kg。另外,去内脏能减少生物胺的生成,在-18、0、4、10、15、20、25和30 ℃贮藏条件下,去内脏后总生物胺最终积累量减少程度分别为14.22%、39.79%、13.83%、29.06%、22.60%、13.56%、26.13%、21.26%。因此,低温冷冻和去内脏处理能够有效控制鲐鱼中生物胺的生成,防止腐败。本研究为有效控制鲐鱼生物胺的产生提供了参考依据。