黄酒酿造过程中产生物胺菌株的筛选及其特性研究

从工业化黄酒酿造环境中筛选具有产生物胺能力的微生物,通过显色反应对菌株的产胺能力进行定性分析,用反相高效液相色谱(RP-HPLC)进行定量分析,获得12株产生物胺的阳性菌株。其中2株菌产生物胺含量较高,经16S r DNA序列分析,菌株AR281为短乳杆菌,菌株AR193为棒状乳杆菌。37℃时,菌株AR281总生物胺的产生量最高为154.76 mg/L;菌株AR193总生物胺的产生量最高为173.78 mg/L。本文研究了发酵温度和发酵时间对其产生物胺能力的影响,为黄酒酿造过程中生物胺含量的控制提供了理论依据。     

休闲发酵色拉米中生物胺的测定与分析

人体摄入过多的生物胺时会出现许多不良反应，本实验对乳杆菌（米酒乳杆菌和戊糖乳杆菌）和片球菌（戊糖片球菌和小片球菌）在休闲发酵色拉米中生物胺的生成量进行了测定与分析。结果表明：供试乳杆菌和片球菌均能产生不等量的组胺、尸胺、腐胺、酪胺、亚精胺和精胺，但都不产生色胺，产品中的生物胺生成量表现出明显的菌株效应。采用复配菌种（片球菌＋葡萄球菌）发酵可以明显降低香肠中组胺、尸胺、腐胺和酪胺以及总生物胺的含量。     

乳酸菌对红曲黄酒酿造过程酵母菌生长及风味物质形成的影响

比较了巴氏葡萄球菌、肠膜明串珠菌、戊糖片球菌、乳酸乳球菌、植物乳杆菌和干酪乳杆菌6种乳酸菌对红曲黄酒酿造体系中酵母菌生长及风味物质形成的影响。结果表明:除干酪乳杆菌外,其它乳酸菌均能促进酵母菌的生长;添加乳酸菌虽然对酿造终产物中还原糖与乙醇含量的影响并不显著,但都能显著提高其有机酸含量。利用顶空固相微萃取-气相色谱-质谱联用技术共鉴定出34种挥发性成分,并对发酵终产物进行感官评定,结果发现正丁酸、苯乙醛、2-苯乙醇、乙酸异戊酯、正丙醇和1-壬醇与感官总分呈正相关,乙酸丁酯、辛酸、乳酸乙酯、癸酸乙酯、辛酸乙酯、正丁醇和5-甲基糖醛与感官总分呈负相关。研究结果表明:在酿造体系中添加植物乳杆菌或干酪乳杆菌均会对红曲黄酒品质产生不利影响,而戊糖片球菌对红曲黄酒品质有一定提升,是潜在可应用于酿造红曲黄酒中的优良乳酸菌菌株。     

一种混合发酵剂对干发酵香肠中生物胺含量的影响

研究了复合发酵剂(戊糖片球菌Pediococcuspentocaceus和木糖葡萄球菌Staphylococcusxylosus)对发酵香肠中生物胺含量的影响。通过与单一发酵剂戊糖片球菌和空白组的产胺、微生物生长情况的比较,分析了木糖葡萄球菌对戊糖片球菌在发酵过程中降胺作用的影响。结果表明,木糖葡萄球菌有助于戊糖片球菌进一步降低发酵香肠中的酪胺和色胺的量,分别比单一发酵菌株降低了6.7%、22.83%;三种胺的量比空白组分别降低了94.04%,29.53%,46.59%,从而证实了戊糖片球菌具有降酪胺和色胺的能力。结合微生物菌数可知,木糖葡萄球菌是通过抑制乳酸杂菌的生长促使戊糖片球菌菌数的增加而有利于胺量的降低。     

发酵香肠中生物胺生成量的菌株效应分析

本实验对乳杆菌(米酒乳杆菌和戊糖乳杆菌)和片球菌(戊糖片球菌和小片球菌)在发酵香肠中生物胺的生成量进行了分析。结果表明,供试乳杆菌和片球菌均能产生不等量的组胺、尸胺、腐胺、酪胺、亚精胺和精胺,但都不产生色胺,产品中的生物胺生成量表现出明显的菌株效应。采用复配菌种(片球菌+葡萄球菌)发酵可以明显降低香肠中组胺、尸胺、腐胺和酪胺以及总生物胺的含量。     

酿酒工艺对黄酒中生物胺的影响

采用高效液相色谱技术,针对黄酒酿造工艺,模拟实际生产,分析了微生物、工艺参数和冷热处理等对黄酒中生物胺含量的影响。结果表明,酵母Y可使生物胺总量降低,霉菌M可使生物胺总量增加,细菌除戊糖片球菌L04使生物胺总量降低外,其他细菌均可使生物胺总量增加;在培养温度32 ℃,培养时间3 d,pH值为3.5,酒精体积分数15%的条件下,生物胺总量为60.19 mg/L;煎酒使生物胺含量增加,冷冻过滤技术因有效降低酪胺含量使生物胺总量大幅降低。     

黄酒发酵过程中乳酸菌的分离及对其产生物胺能力的评价

对黄酒酿造过程中乳酸菌进行分离,并对乳酸菌产生物胺能力进行检测。利用脱羧培养基对分离出的82株乳酸菌进行培养,结合反相高效液相色谱技术(reversed-phase high-performance liquid chromatography,RPHPLC)测定发酵液中生物胺含量,具体色谱条件如下:使用丹磺酰氯(Dansyl Chloride,Dns-CL)进行柱前衍生,以乙腈-乙腈(φ=50%)为流动相梯度洗脱,流速1.0 m L/min,254 nm紫外波检测。该方法在给定浓度范围内线性关系良好(R~20.996),平均回收率为94.46%~105.20%,相对偏差(RSD)均小于5%。检测到组胺产生菌株11株,最大生成量为18.19 mg/L,高产菌株分离自黄酒前酵第2天;酪胺产生菌株28株,最大生成量为30.38mg/L,高产菌株分离自黄酒前酵第2天;腐胺产生菌株51株,最大生成量为299.94 mg/L,高产菌株分离自浸米水。由此表明,有必要控制黄酒发酵过程中的乳酸菌,采用低产或不产生物胺的乳酸菌作为强化菌株,可降低黄酒中的生物胺含量。     

黄酒酿造过程中优势细菌产生物胺的检测与评价

采用显色培养基法、脱羧酶序列特异PCR技术和HPLC分析脱羧酶液体培养基生物胺含量变化3种方法检测和评价来自黄酒酿造过程的41株细菌产生物胺的情况。研究表明,3种方法检测结果大致相同,互补使用能够准确分析细菌产生物胺的情况;41株细菌中,53.7%以上都能产酪胺,31.7%以上产腐胺,24.3%以上产组胺;14株短乳杆菌均携带酪氨酸脱羧酶基因,其中11株菌的酪胺产量超过50.00 mg/L,且多数菌能代谢产生多种生物胺,因此推断其对黄酒中生物胺含量影响最大;细菌代谢是否产生物胺及产生物胺的能力无种属特异性,但有菌株特异性。     

红曲黄酒传统酿造过程中优势细菌活菌检测方法的构建及应用

PMA结合qPCR技术的分子检测手段是一种用于检测活性微生物的有效方法。以红曲黄酒传统酿造体系中的6种优势细菌:巴氏葡萄球菌、肠膜明串珠菌、戊糖片球菌、乳酸乳球菌亚种、植物乳杆菌和干酪乳杆菌为研究对象,设计和合成相应的特异性引物用于实时荧光定量PCR检测,并优化PMA处理条件,通过绘制标准曲线构建6种优势细菌的PMA-qPCR定量检测方法。结果显示:6种特异性引物有效可行,最优的PMA处理条件为PMA处理浓度50μmol/L,交联曝光时间5 min。将构建的PMA-qPCR活菌检测方法应用于红曲黄酒传统酿造过程,发现戊糖片球菌、肠膜明串珠菌和乳酸乳球菌乳亚种是传统酿造体系中最主要的3种细菌。本研究构建的PMA-qPCR活菌检测方法可用于实时定量检测红曲黄酒传统酿造过程中优势细菌及其动态变化,阐明不同细菌在红曲黄酒酿造过程的作用机制。     

熏马肠成熟过程中产生物胺氧化酶菌对生物胺作用的研究

该研究以从新疆熏马肠中筛选出来的6株产生物胺氧化酶菌株(鼠李糖乳杆菌、枯草芽孢杆、腐生葡萄球菌、木糖葡萄球菌、戊糖片球菌、植物乳杆菌)作为发酵剂加入熏马肠中,采用PCR-DGGE技术监测菌落动态变化,高效液相色谱法检测熏马肠中生物胺(色胺、腐胺、苯乙胺、组胺、酪胺、尸胺、精胺、亚精胺)含量变化。结果表明,在熏马肠成熟过程中发酵剂对生物胺有一定的抑制作用。其中以腐生葡萄球菌对色胺、腐生葡萄球菌对苯乙胺、木糖葡萄球菌对尸胺、鼠李糖乳杆菌对腐胺、腐生葡萄球菌对组胺和枯草芽孢杆菌对酪胺的抑制作用最为显著,减少程度分别为45.49%、57.54%、40.75%、51.03%、49.28%、64.33%。本研究为熏马肠以及其他发酵肉制品的提高安全性奠定了基础。     

黄酒生物胺的研究进展

黄酒是我国的传统酿造酒,因其含有丰富的营养物质及独特的风味备受青睐。黄酒发酵过程复杂难控,易产生生物胺等潜在有害物质。生物胺是一类低分子质量含氮有机化合物,过量摄入则会危害人体,对神经、心血管系统造成损伤,产生头痛、心悸、呼吸紊乱、血压变化、呕吐和腹泻等不良症状,严重时还会危及生命。该文对黄酒中生物胺合成、代谢途径及含量水平、影响生物胺形成因素、控制措施及检测方法等方面进行综述,旨在为控制黄酒生物胺含量及提高黄酒质量及安全性提供科学参考。     

Effects of starter cultures and nitrite levels on formation of biogenic amines in sucuk

The effects of starter cultures, starter A (Lactobacillus sakei+Staphylococcus carnosus) and starter B (Pediococcus acidilactici+Staphylococcus xylosus+Lactobacillus curvatus), nitrite levels (0, 75, and 150ppm) and ripening period on biogenic amine production were investigated in sucuk (Turkish dry-fermented sausage) production. Changes in biogenic amines, pH, water activity, moisture, non-protein nitrogen, nitrite and counts of lactic acid bacteria, Micrococcus/Staphylococcus, Enterobacteriaceae, and moulds-yeasts were determined. Ripening period had a very significant effect (P<0.01) on the all biogenic amines. Sausages produced by fermentation with starter cultures, as compared to natural fermentation (control), had lower amounts of putrescine, cadaverine and tyramine (P<0.05). Starter cultures A and B were found to have almost the same effect on the formation of biogenic amines. Use of nitrite in sucuk production affected the formation of biogenic amines except for spermidine and spermine (P<0.01). Lactic acid bacteria counts in the control group were lower than that of starter culture groups. Lactic acid bacteria, Micrococcus/Staphylococcus, and moulds-yeasts counts increased during ripening period while levels of species of Enterobacteriaceae decreased, which was also found to be under the detectable level (<100cfug(-1)), by day 3 of ripening.     

不同乳酸菌发酵剂对发酵红肠品质的影响

为研究乳酸菌发酵对红肠品质的影响,将发酵技术应用于本无发酵工艺的红肠制品中,筛选出能够提高红肠品质的乳酸菌发酵剂。分别将常应用于发酵肉制品的7 种商业乳酸菌发酵剂（木糖葡萄球菌＋戊糖片球菌（THM-17）、木糖葡萄球菌＋清酒乳杆菌＋类植物乳杆菌（PRO-MIX5）、木糖葡萄球菌＋肉葡萄球菌＋清酒乳杆菌（WBL-45）、木糖葡萄球菌＋戊糖片球菌＋植物乳杆菌（VHI-41）、木糖葡萄球菌＋戊糖片球菌＋植物乳杆菌（SHI-59）、肉葡萄球菌＋木糖葡萄球菌（WBX-43）和戊糖片球菌＋木糖葡萄球菌＋肉葡萄球菌＋乳酸片球菌（VBM-60））及8 种单菌（弯曲乳杆菌、戊糖乳杆菌、清酒乳杆菌-1、戊糖片球菌、木糖葡萄球菌、肉葡萄球菌、清酒乳杆菌-2、植物乳杆菌）以107 CFU/g的接种量接种至腌制后的肉馅中,拌馅灌肠后于35 ℃、80%湿度条件下发酵12 h,取样测定发酵后样品的乳酸菌数和细菌总数,再经干燥、蒸煮、烟熏、烘烤制得成品,测定其感官、pH值、色差、质构、亚硝酸盐、硝酸盐、生物胺及N-亚硝胺含量等指标。结果表明：15 种发酵剂中以木糖葡萄球菌和植物乳杆菌2 种乳酸菌发酵剂应用效果较好,所制得产品pH值分别为5.26和5.04,色泽美观,弹性适中,亚硝酸盐残留量（10.84、10.13 mg/kg）低,可显著抑制N-亚硝胺的形成（N-二甲基亚硝胺含量分别为1.29、2.51 μg/kg）,生物胺总量较低。由此说明,木糖葡萄球菌和植物乳杆菌能够显著提高红肠产品的安全品质。     

The influence of lactic acid fermentation on biogenic amines and volatile compounds formation in flaxseed and the effect of flaxseed sourdough on the quality of wheat bread

The aim of the study was to investigate if differences in acidification, biogenic amines (BA) and volatile compounds (VC) occurred when Lactobacillus sakei KTU05-6, Pediococcus acidilactici KTU05-07 and Pediococcus pentosaceus KTU05-8 are used as starters in solid state fermentation of flaxseed and the impact of flaxseed sourdough on the wheat bread making process assessed.     

The effects of a starter culture on the formation of biogenic amines in Turkish soudjoucks

The effects of starter culture containing Lactobacillus sake, Pediococcus pentosaceus, Staphylococcus carnosus plus Staphylococcus xylosus on the formation of biogenic amines during ripening of Turkish soudjoucks were investigated. Determination of eight different biogenic amines was carried out by reverse-phase high performance liquid chromatography (HPLC) using diode array detection. It was found that aerobic plate counts (APC) decreased in the samples with added starter culture. Lactic acid bacteria counts increased in both the controls and samples with added starter during ripening. Total Enterobacteriaceae (ENT) counts decreased on the 30th day for vacuum packed and starter added samples. Yeast counts decreased only in vacuum packed and starter added samples on the 30th day. No biogenic amines were found in ground meat samples. Putrescine (PU) and tyramine (TYR) were determined in all control samples during the storage at 4°C. Starter addition inhibited formation of PU but not TYR.     

Dry sausage fermented by Lactobacillus rhamnosus strains

The ability of three probiotic Lactobacillus rhamnosus strains GG, E-97800 and LC-705 and one commercial Pediococcus pentosaceus starter strain (control) to produce dry sausage was studied. During the fermentation process the numbers of inoculated lactic acid bacteria increased from approx. 7 log10 to 8-9 log10 cfu/g and the pH values decreased from 5.6 to 4.9-5.0. The sensory test indicated that the dry sausages fermented by L. rhamnosus LC-705 were inferior to the control sausages. The presence of inoculated experimental strains as predominant organisms in the dry sausages was recognised on the basis of their genetic fingerprints by ribotyping. The concentrations of biogenic amines remained low during the ripening process. These results indicated that the studied Lactobacillus rhamnosus strains, especially strains GG and E-97800, are suitable for use as probiotic starter cultures in fermenting dry sausage.     

葡萄酒中生物胺含量的影响因素研究

研究了葡萄酒生产过程中影响生物胺产生量的主要因素,结果表明,葡萄酒酒精发酵和苹果酸．乳酸发酵过程均有牛物胺的产生,影响葡萄酒中生物胺产生的主要因素是乳酸菌中氨基酸脱羧酶的活性,本研究添加了氨基酸脱羧酶辅酶的样品与对照相比,生物胺含量提高了90．6％。酵母菌和乳酸菌种类、酒精发酵温度,生物胺前体物质也是影响生物胺含量的主要因素,发酵温度越高、生物胺产生量越;在一定范围内,氨基酸的浓度越大产生物胺越多。但酵母菌和乳酸菌的添加量对生物胺浓度的影响较小。