产胺氧化酶发酵剂对新疆熏马肠中生物胺影响及其作用机制

Smoked horsemeat sausage is one form of spontaneously fermented sausages which is appreciated by the consuming public as peculiar flavour. Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in fermented sausage. The amine oxidase-production bacterias isolated in our lab will be used as strarter in this project. After amine oxidase is separated and extracted, characteristic of the oxidase will be studied. The effect of different amine oxidase-production bacteria and different biogenic amines on expression of amine oxidase gene and phenotypic plasticity will be studied by Real time-PCR and HPLC technology. In order to determine the information transfer mode between amine oxidase-production bacteria, signaling molecule will be detected by LC-MS. Then mechanism amine oxidase generation was found. After the effect of adjuvant material and smoke element on oxidase-productive activity and the enzyme activity was analysed , the influence mechanism of biogenic amines oxidation will be established during smoked horsemeat prosessing. In order to find competition relations between starter and biogenic amines-productive bacteria in smoked horsemeat sausage, the change of bacteria protortion and the content of biogenic amines will be study by HPLC and PCR-DGGE technology. After the effects on production of amine oxidase and enzyme activity was researched systematically, the effect of starter on biogenic content in smoked horsemeat sausage and the mechanism will be explained clearly on molecular level.

新疆熏马肠是一种自然发酵香肠，因风味独特深受消费者喜爱，但生物胺含量较高，存在安全隐患。以产胺氧化酶微生物作发酵剂是减少熏马肠中生物胺累积的重要途径。利用已分离到的产胺氧化酶菌株作发酵剂，从中分离提取胺氧化酶并研究其特性；采用Real time-PCR，HPLC方法研究不同微生物之间、不同生物胺底物对产胺氧化酶基因表达和发酵剂表型可塑性的影响，LC-MS检测群体感应信号分子及研究信息传递方式，揭示胺氧化酶产生机制；研究熏马肠加工中常用辅料、烟熏成份对发酵剂产酶能力和酶活的影响，明确熏马肠加工环境因素对生物胺氧化影响机制；采用PCR-DGGE方法研究熏马肠生产过程中菌相变化，HPLC方法检测生物胺的动态变化，从而明确发酵剂繁殖与内源产生物胺微生物竞争关系及对生物胺影响。本项目针对影响发酵剂产酶和酶活因素进行系统研究，从分子水平阐明产胺氧化酶发酵剂在熏马肠中对生物胺影响及其作用机制。

新疆熏马肠是一种自然发酵香肠，因风味独特深受消费者喜爱，但生物胺含量较高，存在安全隐患。以产胺氧化酶微生物作发酵剂是减少熏马肠中生物胺累积的重要途径。分离出产生物胺氧化酶并符合肉品发酵条件的菌株6株。研究了产生物胺氧化酶菌的酶学特性、不同产生物胺氧化酶菌之间对氧化酶产生的影响、不同环境条件对产酶的影响以及不明确了前体物质对菌株表型特征的影响。结果表明，生物胺浓度从100-600μg/ml时，随着生物胺浓度的增加，菌对株生物胺的降解能力和基因表达能力都逐渐减弱。当NaCl浓度达到8%时，乙醇浓度达到4.5%时，亚硝酸盐达到4.0g/kg微生物对生物胺的降解活性被强烈抑制。烟熏产生物胺氧化酶发酵剂生长和产酶都没有显著影响。不同产生物氧菌株之间没有显著的协同作用，但可能有抑制作用，合理搭配很重要。本项目的研究，为提高熏马肠等发酵肉制品安全性提供保障。