黄酒中生物胺的形成及其影响因素

采用高效液相色谱技术,分析了黄酒酿造原料、发酵剂和不同年份酒中的生物胺含量,并对传统工艺和机械化生产过程中生物胺含量的变化进行了跟踪分析,剖析了影响生物胺含量的因素。结果表明,腐胺和酪胺是黄酒中的主要生物胺,糯米原料和曲中生物胺含量很低,检出的3种脂肪族生物胺(腐胺、尸胺和精胺)总量不超过6.01mg/kg;7种酒母中生物胺总含量差异极显著(P<0.01),总量变化范围为16.43～87.72 mg/L;酒母中生物胺总量与总酸呈正相关,与酒精度和感官品质呈显著性负相关;机械化工艺和传统工艺发酵过程中生物胺均呈先升后降的变化趋势,但传统工艺比机械化工艺生物胺含量高;发酵过程中氨基酸逐渐增多,与生物胺含量变化无相关性;发酵醪中细菌总数在前期急剧增加到最大值,之后逐渐降低,变化趋势与生物胺含量变化相同;陈酿时间对传统工艺生物胺含量影响明显,随着时间延长,生物胺含量降低,但陈酿时间对机械化工艺无明显影响。本实验检测的黄酒发酵醪及年份酒中生物胺总量范围为0.14～175.15 mg/L,不存在安全问题。     

黄酒前酵中生物胺生成规律的研究

本研究对黄酒前酵工序的生物胺生成规律及影响因素进行了分析探讨。采用反相高效液相色谱技术,改进了生物胺定量检测法,该检测体系准确可靠,样品峰型对称分离度好且缩短了检测时间。分析了前酵工序中主要微生物,氨基酸,发酵醪酸度、糖度、酒精度、p H对生物胺生成的影响。发现前酵工序对成品酒生物胺的影响度为77.67%,第一次开耙阶段生物胺总量增幅最大,达到7.63 mg/L。研究发现主要酿造微生物中乳酸菌总数与生物胺总量成正相关,乳酸菌在搭窝期间生长速率最大,为7.13×106CFU/(m L·h)。在前酵工序中生物胺总量与发酵醪酸度、酒精度、p H呈显著正相关,与糖度呈负相关;前酵过程中主要生物胺与前体氨基酸也呈明显正相关。本研究分析了黄酒前酵中生物胺生成规律,有助于建立降低黄酒生物胺含量的更安全、科学的工艺。     

Reduction of biogenic amine formation using a negative amino acid-decarboxylase starter culture for fermentation of Fuet sausages

The ability of Lactobacillus sakei CTC494, a negative amino acid-decarboxylase starter culture, to reduce biogenic amine accumulation during sausage fermentation and storage at 4 and 19 degrees C was studied. The effect on the amine formation of the tyramine producer Lactobacillus curvatus CTC371, as a positive strain, was also examined in comparison to a spontaneous fermentation process without starter culture (control batch). The polyamines spermine, spermidine, and diaminopropane were not influenced by the ripening, and their levels slightly decreased in all the batches throughout the storage. Tyramine, cadaverine, and putrescine were the main amines formed during the ripening. The addition of starter culture resulted in a decrease on the biogenic amine formation, depending on the strain inoculated. A great reduction in tyramine content was achieved when L. sakei CTC494 was inoculated, whereas L. curvatus CTC371 only attenuated tyramine accumulation compared with the control batch. Both starters were able to significantly limit the production of putrescine and cadaverine, and they inhibited tryptamine and phenylethylamine formation by the wild microbial flora. Tyramine levels of the control sausages rose during the storage at both temperatures, whereas those of cadaverine only increased at 19 degrees C. On the contrary, sausages manufactured through the starter controlled fermentation did not show changes of amine contents during the storage. The addition of a proper selected starter culture is advisable to produce safer sausages with low contents of biogenic amines.     

Biogenic Amine Content of Beers Consumed in Turkey and Influence of Storage Conditions on Biogenic Amine Formation

The biogenic amine content of beers available in Turkey, both domestic production and imported products, was investigated. A total of 17 domestic and 13 imported beers were evaluated in terms of 8 different biogenic amines (histamine, tyramine, spermine, spermidine, 2‐phenylethylamine, putrescine, tryptamine and cadaverine). HPLC methodology with pre‐column derivatization and photodiode array detection after derivatization with dansyl chloride was used for quantification. In addition, the evolution of important amines such as histamine, tyramine, putrescine and cadaverine were investigated during different storage conditions by response surface methodology. The results indicated that both storage temperature and storage time were important factors affecting biogenic amine content. Histamine and putrescine increased during storage, but then decreased after reaching a maximum level after six weeks. With the biogenic amines tyramine and cadaverine, the amounts increased during the entire storage period. At higher storage temperatures, the formation of biogenic amines increased.     

Effectiveness of a Lactobacillus sakei starter culture in the reduction of biogenic amine accumulation as a function of the raw material quality

The effectiveness of an amine-negative starter culture (Lactobacillus sakei CTC494) in the reduction of biogenic amine production during the ripening of fermented sausages was examined. Four batches were manufactured in parallel: spontaneously fermented and starter-mediated sausages were manufactured from two lots of raw materials of different hygienic quality. Besides the biogenic amine contents, changes in the microbial counts, nitrogenous fractions, pH, and water content were measured at several sampling points during the ripening process. In sausages manufactured from good quality meat, the starter strain of L. sakei reduced and even inhibited biogenic amine accumulation during sausage fermentation, the end products showing extremely low biogenic amine contents (tyramine levels less than 15 mg/kg of dry matter and putrescine and cadaverine levels less than 5 mg/kg of dry matter). Nevertheless, starter-mediated sausages made from poorer-quality raw materials showed much higher amine contents (308, 223, and 36 mg/kg of dry matter of cadaverine, tyramine, and putrescine, respectively), which were only slightly lower than those of the spontaneously fermented sausages made from the same raw materials. The relatively high bacterial numbers of raw materials of poorer-hygienic quality diminished the beneficial effect of the starter strain. Therefore, the effectiveness of the starter was strongly dependent on the hygienic quality of the raw materials used.     

Ferulic Acid Release and 4‐Vinylguaiacol Formation during Chinese Rice Wine Brewing and Fermentation

The release of ferulic acid (FA) and the subsequent enzymatic decarboxylation to 4‐vinylguaiacol (4‐VG) were nearly ubiquitous during the production of Chinese rice wine. To evaluate the release of FA and the transformation to 4‐VG, the levels of FA and 4‐VG were determined by high performance liquid chromatography (HPLC) and gas chromatography‐mass spectrum (GC‐MS), respectively. During rice wine brewing, the concentrations of FA and 4‐VG increased as fermentation time prolonged, with a rapid rise in days 1 to 5 of the main fermentation and followed by a slower rise from the 6th day post inoculation. FA and 4‐VG levels in mashes, fermented by yeast with wheat Qu, were significantly higher than those in mashes fermented by yeast using commercial enzyme additions. The release of FA from rice by wheat Qu involves an enzymatic release mechanism. The levels of FA and 4‐VG from rice flour by wheat Qu rose gradually during the first 24 h and then tended to stabilize. The capacity of yeasts to decarboxylate phenolic acids (Pof⁺ phenotype) was absent in most rice wine brewing yeasts. These results suggest that the production of FA and 4‐VG originated from materials in the wheat Qu rather than by the yeast during Chinese rice wine brewing and fermentation.     

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。     

我国黄酒研究现状与发展趋势

传统黄酒酿造是利用酒药(小曲),酒曲(麦曲、红曲)中多种有益微生物参于糖化发酵。由多种霉菌,酵母菌和细菌等共同作用及发酵过程中产生的代谢产物构成了黄酒特有的色,香,味,格。该文对黄酒酿造过程的麦曲功能,淋饭酒母,主酵,后酵工艺及黄酒中风味形成等的相关研究进行综述与总结,并对今后的研究方向进行了探讨。     

Wheat Qu and Its Production Technology,Microbiota, Flavor,and Metabolites

Chinese rice wine (CRW) is an ancient wine with a moderate alcohol content; it has been prized in China for its color, sweet flavor profile, and is a national beverage typically used in banquets. Wheat Qu is the starter used to make CRW, and plays a vital role in the quality and flavor profile. This review details the raw materials and the wheat Qu manufacturing process, and recent information on wheat Qu microbiotas. The volatile compounds in wheat Qus and its flavor are also reviewed. Finally, the enzymes, and metabolites contained in wheat Qu, such as biogenic amines, are also discussed.     

不同发酵方式对发酵马肉肠贮藏期品质劣变及食用安全性的影响

为有效确保发酵马肉肠贮藏期间的品质及安全性,研究添加汉逊德巴利酵母菌1808（Debaryomyces hansenii 1808）、木糖葡萄球菌21445（Staphylococcus xylose 21445）、植物乳杆菌E11（Lactobacillus plantarum E11）的复合发酵剂发酵组（FP）和单一乳酸菌发酵组（DZ）、自然发酵组（ZR）马肉肠贮藏过程中生物胺的变化,考察不同发酵方式对发酵马肉肠pH值、水分活度（aw）、色差、总挥发性盐基氮（total volatile basic nitrogen,TVB-N）含量、硫代巴比妥酸反应物（thiobarbituric acid reactive substances,TBARs）值及菌落总数的影响。结果表明：在贮藏期间,相比于自然发酵组和单一乳酸菌发酵组,添加复配发酵剂可有效减缓发酵马肉肠TVB-N含量、TBARs值、pH值的增幅,抑制贮藏后期有害杂菌的生长,防止产品过早发生腐败变质;此外,复配发酵剂可显著降低贮藏过程中马肉肠的水分损失（P＜0.05）,维持色泽的稳定,抑制生物胺（尸胺、腐胺、酪胺、组胺）的积累。综上,随着贮藏时间的延长,复配发酵剂可显著抑制发酵马肉肠品质的劣变及不良生物胺的过多积累。     

Formation of biogenic amines throughout the industrial manufacture of red wine

Changes in biogenic amines (histamine, methylamine, ethylamine, tyramine, phenylethylamine, putrescine, and cadaverine) were monitored during the industrial manufacture of 55 batches of red wine. The origin of these amines in relation to must, alcoholic fermentation, malolactic fermentation, sulfur dioxide addition, and wine aging and the interactions between amines and their corresponding amino acids and pH were statistically evaluated in samples from the same batches throughout the elaboration process. Some amines can be produced in the grape or the musts (e.g., putrescine, cadaverine, and phenylethylamine) or can be formed by yeast during alcoholic fermentation (e.g., ethylamine and phenylethylamine), although quantitatively only very low concentrations are reached in these stages (less than 3 mg/liter). Malolactic fermentation was the main mechanism of biogenic amine formation, especially of histamine, tyramine, and putrescine. During this stage, the increase in these amines was accompanied by a significant decline in their amino acid precursors. Significant correlations between biogenic amine formation and the disappearance of their corresponding amino acids were observed, which clearly supports the hypothesis that malolactic bacteria are responsible for accumulation of these amines in wines. No increase in the concentration of biogenic amines was observed after SO2 addition and during wine aging, indicating that sulfur dioxide prevents amine formation in subsequent stages.     

发酵剂对牦牛乳硬质干酪成熟过程中生物胺的影响

乳酸菌产生物胺的能力具有菌株特异性,因此,为了探究不同种类发酵剂对牦牛乳硬质干酪中生物胺形成的影响,该试验利用高效液相色谱对3种不同发酵剂制作的硬质干酪成熟过程中生物胺进行了测定和分析。结果表明,嗜热和嗜温发酵剂牦牛乳硬质干酪中检测出2-苯乙胺、腐胺、尸胺、组胺和酪胺,混合发酵剂干酪中检测出腐胺、2-苯乙胺、尸胺和酪胺。各生物胺之间呈现正相关性。3种不同发酵剂干酪在1～6个月成熟过程中,其各生物胺整体呈现增加趋势,嗜热、嗜温和混合发酵剂干酪中总生物胺最高含量分别为(448.3±9.6)、(456.8±58.4)、(293±24.5)mg/kg。组胺和酪胺是2种毒性相对高的生物胺,嗜热发酵剂干酪中组胺和嗜温发酵剂干酪中酪胺最高,其最高含量分别为(20.8±7.9)、(92.9±6.7)mg/kg,混合发酵干酪中未检测出组胺,酪胺含量次之,3种不同发酵剂干酪中组胺、酪胺含量均低于推荐安全剂量50 mg/kg和100 mg/kg。这为合理选择发酵剂和控制干酪中生物胺形成提供了依据。     

利用高效液相色谱法测定不同发酵时长郫县豆瓣酱中的8种生物胺

本研究基于高效液相色谱(HPLC)技术,测定了不同发酵时长(1~96个月)郫县豆瓣酱中色胺、β-苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺8种生物胺的含量。结果显示,HPLC分析中,8种生物胺的检出限在0.17~0.35 mg/L之间,定量限在0.55~0.92 mg/L之间,各生物胺标品在1~60 mg/L的质量浓度范围内线性关系良好,可被准确定量;各生物胺回收率在73.10%~104.13%之间,说明本方法具有较高的重复性和灵敏度。腐胺、尸胺、酪胺和组胺是郫县豆瓣酱中的主要生物胺;随着发酵时间的延长,腐胺含量快速上升,尸胺含量在发酵前54个月迅速上升,之后明显下降,酪胺和组胺在发酵54个月后含量有所增加;其他生物胺含量较低,且发酵期间变化不明显;郫县豆瓣酱中总生物胺含量随发酵时间的延长不断积累,从(37.86±6.69) mg/kg增加到(155.09±8.53) mg/kg,其含量在发酵前54个月快速上升,之后趋于稳定。目前食品中的多数生物胺尚没有限量标准,但参考现有的毒理学数据来看,本研究中郫县豆瓣酱的生物胺含量水平相对安全。     

Changes in Volatile Compounds of Chinese Rice Wine Wheat Qu During Fermentation and Storage

In order to understand the optimum time of using wheat Qu for Chinese rice wine brewing on the basis of the contents of volatile compounds, changes in volatile compounds of Chinese rice wine wheat Qu during fermentation and storage were studied by headspace solid phase microextraction (HS‐SPME) combined with gas chromatography‐mass spectrum (GC‐MS). It was shown that the concentrations of most groups of volatiles, including alcohols, aldehydes, aromatic compounds, phenols, sulphides, furans, and nitrogen‐containing compounds, increased progressively from the first day to the fourth day of fermentation, and then decreased gradually. A minority group of volatile compounds such as acids and ketones peaked after one day of fermentation, esters increased slowly from the first day to the eighth day of fermentation. Lactones and esters rose slowly from the first day to the fifteenth day of fermentation respectively, and decreased afterwards, whereas the evolution of terpenes was somewhat erratic. These results suggested that changes in the concentrations of most groups of volatile compounds during fermentation and storage were uniform. This made it possible to determine the optimum level of fermentation and storage for Chinese rice wine wheat Qu on the basis of the contents of volatile compounds.     

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。     

北工商海洋杆菌诱导戊糖片球菌产生细菌素及降黄酒生物胺功效研究

利用功能微生物控制杂菌污染,是黄酒生产中控制生物胺含量的有效途径。从黄酒麦曲中分离筛选出新菌种[北工商海洋杆菌(Pontibacter beigongshangensis)]和新菌株[戊糖片球菌(Pediococcus pentosaceus)M25]。北工商海洋杆菌能够诱导戊糖片球菌产生细菌素,对黄酒中产生物胺的腐生葡萄球菌(Staphylococcus saprophyticus)、阪崎肠杆菌(Cronobacter sakazakii)、乳酪短杆菌(Brevibacterium casei)、芽孢杆菌(Bacillus)和阴沟肠杆菌(Enterobacter cloacae)均具有抑制作用。北工商海洋杆菌和戊糖片球菌自身不产生物胺。添加北工商海洋杆菌和戊糖片球菌,制备强化功能曲,利用强化功能曲酿造黄酒,可使黄酒中的生物胺含量降低29.8%,而对黄酒的风味物质影响不明显。因此,未来北工商海洋杆菌和戊糖片球菌可以作为降生物胺功能菌株应用在黄酒酿造中。     

豆豉中生物胺含量的动态变化研究

采用高效液相色谱法对豆豉中生物胺含量进行动态检测。结果表明,纯种强化发酵和自然发酵豆豉样品中均检测出色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺。纯种强化发酵条件下生物胺总含量在前、后发酵时期都呈先上升后下降的趋势,而自然发酵条件下均呈缓慢上升趋势,且前发酵总生物胺含量高于后发酵时期。两种发酵条件下,酪胺和亚精胺为主要生物胺,苯乙胺存在纯种发酵中。纯种强化发酵豆豉总生物胺含量高于自然发酵,酪胺和苯乙胺的含量分别高出2倍和4倍,与生产环境中粪肠球菌的污染关联性较大。豆豉发酵过程和成品中总生物胺含量均＜250 mg/kg,短期内不存在食品安全问题。     

青稞酒发酵过程中生物胺动态变化

对青稞酒发酵过程中生物胺的动态变化规律进行研究。利用氨基酸自动分析液相色谱法测定青稞酒不同发酵阶段中7种生物胺的含量,来评价发酵环境以及控制合理发酵过程和发酵终产物的质量。结果表明：青稞原料中不含组胺,但是青稞酒醅及成品青稞酒中都含有组胺。传统发酵和多菌种发酵两种发酵方式青稞酒的生物胺含量变化规律是：腐胺及胍丁胺的含量变化随着发酵的进行呈下降趋势;组胺及酪胺的含量随发酵进程逐渐上升,尤其是酪胺,上升趋势明显,多菌种发酵由2.98μg/g上升至5.36μg/g,传统发酵由5.97μg/g 上升至11.67μg/g。其余各种生物胺含量变化不明显,发酵过程的生物胺总量变化也不显著。本实验检测的青稞酒醅中的生物胺总量范围在53.44～72.56μg/g,不存在生物胺安全问题。     

大蒜精油和发酵剂对熏马肠中生物胺及微生物分布的影响

为研究大蒜精油和发酵剂对熏马肠中生物胺的影响,采用高效液相色谱法测定熏马肠中生物胺含量,利用 宏基因组学技术分析熏马肠中微生物多样性和生物胺含量的变化情况。结果表明：将大蒜精油和发酵剂联合使用能 够很好地减少熏马肠中尸胺、腐胺、组胺、酪胺和苯乙胺的产生,对控制生物胺在熏马肠中的含量起到非常显著的 作用;熏马肠中含有丰富的微生物物种,通过样品科分类水平物种分析,得出乳酸菌在熏马肠整个加工过程都是优 势菌种,而葡萄球菌在成熟后期数量较多。     

猪肉自然发酵香肠中细菌和乳酸菌对生物胺积累的作用

在猪肉香肠的自然发酵时，检测了选择的污染菌对形成的生物胺的性质和产量的作用。本文主要研究的是在肉腐败过程中胺的变化和微生物生长的变化，而且重点研究的是加入产酪胺的短乳杆菌（L．brevis）和产尸胺的阴沟肠杆菌（E．cloacae）的猪肉发酵香肠在发酵时生物胺的积累。在腐败和香肠发酵时相关微生物和胺的量较多。尸胺的高度积累和大量的不理想肠细菌有关，在香肠发酵时，酪胺的产生并不完全是乳酸菌的作用。另一方面，也涉及到必要的生产香肠的自然发酵菌，污染菌对于腐胺和其它生物胺的作用，现在还不清楚。