CN102106373A - 一种德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用 - Google Patents
一种德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用 Download PDFInfo
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Abstract
本发明名称为一种德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用，适用于冷却生鲜猪肉馅的防腐。本发明的德氏乳杆菌乳酸亚种(Lactobacillusdelbrueckii subsp.lactis)zhang-LF CGMCC No.4393分离筛选自福建农贸市场中的干鱼制品，其产生的细菌素可被人体内蛋白酶所降解。本发明德氏乳杆菌乳酸亚种细菌素应用于冷却猪肉馅防腐，其防腐手段简单，细菌素用量少，成本低，适用于规模化冷却猪肉馅防腐。
Description
技术领域
本发明涉及食品领域中一种德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用。
背景技术
肉制品由于自身内在的环境条件，如pH、水分活度和营养成分等，容易滋生微生物，鲜肉中常见的病原性细菌有单核细胞增生李斯特菌(简称单增李斯特菌)、沙门氏菌、肠球菌、布鲁氏杆菌、炭疽杆菌、猪丹毒丝菌、金黄色葡萄球菌、肉毒梭菌、产气荚膜梭菌、溶血性链球菌和小肠结肠炎耶森氏菌等。目前猪肉中控制单增李斯特菌的生长是一个较为棘手的问题，食入单增李斯特菌侵染的猪肉会导致小部分人群感染，尤其孕妇、婴儿和免疫力低下的人群，在这些被感染的人群中致死率是偏高的，在27％～34％之间。单增李斯特菌是肉制品中广泛存在的腐败菌，它耐低温，故冷藏对它生长的抑制作用不大，该菌在有氧和无氧条件下都能生长，故真空包装或二氧化碳气调保藏都不能防止该腐败菌的生长。此外，pH、氯化钠、硝酸盐对其生长的影响也不明显。乳酸菌会伴随肉制品的加工过程而生长繁殖，乳酸菌可通过几种途径来抑制一些腐败菌的生长，比如代谢产生乙酸或乳酸，会导致环境pH降低，由此抑制不耐酸细菌的生长；其次乳酸菌能产生具有抑菌效果的代谢产物，即细菌素。细菌素对人和动物无毒性、无抗原性、对热稳定，易被人体消化道的一些蛋白酶所降解，故不会在人体中过度积累而引起不良反应。随着消费者对化学合成防腐剂添加量的关注，以及对天然食品防腐剂的需求，国内外学者们逐渐将研究方向转移到这些既可抑菌且无毒物残留的乳酸菌细菌素的开发应用上来，期望将乳酸菌细菌素添加到肉制品中，以控制腐败菌的生长。
乳酸菌细菌素是乳酸菌代谢过程中通过核糖体合成机制产生的一类具有抑菌活性的多肽或蛋白质类物质，抑菌对像主要是近缘菌株。虽然国内外的研究者们对于天然抑菌物质细菌素的筛选正进行着不断的探索，但只有Nisin是世界上公认天然的食品防腐剂，是至今唯一允许用作食品防腐剂的乳酸菌细菌素，到目前为止，已在世界约50个国家和地区得到广泛应用。但是，Nisin的抑菌谱较窄，pH8.0以上即失活，只能在酸性食品中发挥作用，Nisin主要抑制革兰氏阳性菌，对革兰氏阴性菌的抑菌作用不理想。相对而言，Nisin在乳制品中的抗菌作用要优于在肉制品中的作用，故需要筛选适用于肉制品防腐保鲜的细菌素。
目前，国内文献中有关乳酸菌细菌素作为防腐剂在肉制品中应用的报道：Pediocin PA-1对肉制品中的腐败菌如单增李斯特菌、弯曲乳杆菌具有较好的抑制作用，Pediocin PA-1比较适用于鸡肉中抑制单增李斯特菌；Rosalinda Urso等在自然发酵香肠中筛选到一株沙克乳杆菌Lacticbacteria sakei，该菌株能产细菌素Sakacin P，此细菌素不但能较好地抑制香肠中的肠球菌，而且对香肠的风味无不良影响。Samier Ananou等分别将纯的或半纯的细菌素Enterocin AS-48和产细菌素菌株Enterococcus f aeciumA-48-32接种到香肠中，两者都能较好地抑制金黄色葡萄球菌的生长。国内专利中有关乳酸菌细菌素在肉制品中应用的报道：南京农业大学申请的“一种产细菌素的米酒乳杆菌菌株及其用法”，专利申请号为：CN03132323.5；中国农业大学申请的“一种戊糖乳杆菌细菌素及其专用生产菌株与应用”，专利申请号为：CN200510105985.5。德氏乳杆菌乳酸亚种(Lactobacillus delbrueckii subsp.Lactis)是中华人民共和国卫生部办公厅于2010年4月22日印发的《可用于食品的菌种名单》中明文规定的菌种之一(见附件)，因此，为德氏乳杆菌乳酸亚种的细菌素作为食品防腐剂提供了可靠依据。目前国内发明专利中尚无关于德氏乳杆菌乳酸亚种细菌素作为防腐剂在肉制品中的应用报道。
发明内容
本发明的目的是提供一种德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用。德氏乳杆菌乳酸亚种(Lactobacillus delbrueckii subsp.lactis)zhang-LF，已于2010年12月1日保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC)，保藏号为：CGMCC No.4393。德氏乳杆菌乳酸亚种细菌素，是从德氏乳杆菌乳酸亚种zhang-LF发酵液中提取的。
本发明确定德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中应用的工艺流程：活猪、致昏、放血、去头、剥皮、摘除内脏、消毒、分割、添加细菌素冻干粉并绞肉、冷却、包装。其中在绞肉的同时添加德氏乳杆菌乳酸亚种细菌素冻干粉，直至搅拌均匀，细菌素效价为1280AU/mL，每公斤猪肉馅添加细菌素冻干粉为1.25g，产品置于4℃保藏。
上述的德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用属于本发明保护范围。
附图说明
德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中应用的工艺流程图说明如下：
(1)活猪：检验检疫合格；(2)致昏：电压85V，时间3～4s。(3)放血：采用吊挂方式，时间4min；(4)去头、剥皮、摘除内脏；(5)消毒：脱氢乙酸(0.001％)，5s；(6)分割、绞肉：添加德氏乳杆菌乳酸亚种细菌素冻干粉与绞肉同时进行，细菌素效价1280AU/mL，添加量为每公斤猪肉馅添加细菌素冻干粉为1.25g；(7)冷却：0～4℃，10h，风速1m/s；(8)包装：产品置于4℃保藏。
德氏乳杆菌乳酸亚种(Lactobacillus delbrueckii subsp.lactis)zhang-LF分离筛选自福建农贸市场中的干鱼制品，其产生的细菌素可被人体内蛋白酶所降解。本发明德氏乳杆菌乳酸亚种细菌素应用于冷却猪肉馅防腐，其防腐手段简单，细菌素用量少，成本低，适用于规模化冷却猪肉馅防腐。
具体实施方式
下述实施例中的实验方法，如无特别说明，均为常规方法。
下述实施例中的百分含量，如无特别说明，均为体积百分含量。
实施例1、一种德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用
1、德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中的应用
(1)将购买的冷却猪肉馅各称取5g，分装到2个已灭菌的100mL三角瓶中，与灭菌的45mL 0.85％的生理盐水混合，取其中1个三角瓶，加入6.25mg效价为1280AU/ml的德氏乳杆菌乳酸亚种细菌素冻干粉，充分混匀后静置30min，加入500μL的单增李斯特菌液，冷却猪肉馅渗出液中单增李斯特菌初始菌落总数为5.7×103cfu/mL；另1个三角瓶中加入6.25mg加热灭活的德氏乳杆菌乳酸亚种细菌素冻干粉，作为空白对照，充分混匀后静置30min，加入500μL的单增李斯特菌液，冷却猪肉馅渗出液中单增李斯特菌初始菌落总数为5.7×103cfu/mL，将2个三角瓶于37℃静置30min，4℃冷藏10d，每天取样检测；
(2)将2个三角瓶中的混合物在涡旋混匀仪上振荡混匀(回转直径4.5mm，转速为1500转/mm，振荡10s)，各取1mL猪肉馅渗出液，10倍梯度稀释，在无菌条件下取100μL稀释液涂布在李斯特固体培养基上，倒置于37℃温箱中培养，检测单增李斯特菌的菌落总数。每天从试管中取样检测单增李斯特菌的菌落总数(步骤同上)，共10d。
2、德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐应用中的抑菌效果评定
德氏乳杆菌乳酸亚种细菌素添加到冷却猪肉馅后，在10d的保藏期内每天计算试验组和空白对照组中单增李斯特菌的菌落总数，并计算抑菌率。
抑菌率计算公式为：
X＝(A-B)/A×100％
式中X为抑菌率(％)；
A为空白对照组单增李斯特菌的菌落总数(cfu/mL)；
B为试验组单增李斯特菌的菌落总数(cfu/mL)。
评价标准：抑菌率≥50％～90％，细菌素有抑菌作用，抑菌率≥90％，细菌素有较强抑菌作用。
10天的保藏期内该细菌素对单增李斯特菌有较强的抑菌作用，试验结果见表1。
表1德氏乳杆菌乳酸亚种细菌素
在冷却猪肉馅防腐中对单增李斯特菌抑菌作用的试验结果
10天的保藏期内抑菌率均≥50％～90％，说明德氏乳杆菌乳酸亚种细菌素对单增李斯特菌的生长有抑制作用，从第3天开始至第10天结束，抑菌率≥90％，说明德氏乳杆菌乳酸亚种细菌素对单增李斯特菌的生长有较强的抑菌作用，试验结果表明，德氏乳杆菌乳酸亚种细菌素添加到冷却猪肉馅中，至少10d内对食品腐败菌单增李斯特菌的生长有抑制作用，由此可见，德氏乳杆菌乳酸亚种细菌素添加到冷却猪肉馅中具有明显的防腐作用。
上述试验确定德氏乳杆菌乳酸亚种细菌素在冷却猪肉馅防腐中应用的工艺流程：活猪、致昏、放血、去头、剥皮、摘除内脏、消毒、分割、添加细菌素冻干粉并绞肉、冷却、包装。其中：绞肉的同时添加德氏乳杆菌乳酸亚种细菌素冻干粉，直至搅拌均匀，细菌素效价为1280AU/mL，每公斤猪肉馅添加细菌素冻干粉为1.25g，产品置于4℃保藏。
Claims (1)
1.一种利用德氏乳杆菌乳酸亚种(Lactobacillus delbrueckiisubsp.lactis)zhang-LF CGMCC No.4393细菌素在冷却猪肉馅防腐中应用的方法，包括活猪、致昏、放血、去头、剥皮、摘除内脏、消毒、分割、添加细菌素冻干粉并绞肉、冷却、包装，其特征在于：绞肉的同时添加德氏乳杆菌乳酸亚种细菌素冻干粉，直至搅拌均匀，细菌素效价为1280AU/mL，每公斤猪肉馅添加细菌素冻干粉为1.25g，产品保藏温度为4℃。
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| CN101703304A (zh) * | 2009-12-01 | 2010-05-12 | 郑州三全食品股份有限公司 | 一种保鲜剂及其在馅料中的应用 |
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| 《水利渔业》 20071231 郭良辉等 溶菌酶与Nisin符合生物保鲜剂对蚌肉的保鲜效果 112-114 1 第27卷, 第4期 * |
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