CN111411057B - 一株产广谱抗菌肽的棒状乳杆菌及其抗菌肽的应用 - Google Patents
一株产广谱抗菌肽的棒状乳杆菌及其抗菌肽的应用 Download PDFInfo
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- CN111411057B CN111411057B CN202010319752.XA CN202010319752A CN111411057B CN 111411057 B CN111411057 B CN 111411057B CN 202010319752 A CN202010319752 A CN 202010319752A CN 111411057 B CN111411057 B CN 111411057B
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- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
本发明提供一株产广谱抗菌肽的棒状乳杆菌及其抗菌肽的应用,属于微生物技术领域。本发明的棒状乳杆菌(Lactobacillus coryniformis)FZU63,其保藏编号为:CGMCC NO.19553。该菌株产生的抗菌肽对多种革兰氏阳性菌和阴性菌具有显著的抑制作用。棒状乳杆菌FZU63产生的抗菌肽AMP‑fzu63包含序列为RQQPMTLDYRW、LVKNAKERGY的2条多肽序列。棒状乳杆菌FZU63产生的抗菌肽AMP‑fzu63应用于海水鱼保鲜中,能通过抑制鱼肉中微生物的生长与繁殖,从而降低不良气味物质己醛和1‑辛烯‑3‑醇的产生。
Description
技术领域
本发明属于微生物领域,具体涉及一株产广谱抗菌肽的棒状乳杆菌及其抗菌肽的应用。
背景技术
鱼肉的腐败变质通常由自溶酶分解、脂肪氧化和微生物滋生三个因素引起;其中,微生物生长和代谢是导致鱼肉腐败的主要原因。在我国沿海地区盛产各种海水鱼,经捕捞离水上岸后因气压、温度、溶氧等环境因素骤变,大部分海水鱼不易存活。海水鱼死亡后,细胞代谢终止,表皮杯状细胞停止分泌含有抗菌物质的黏液;此外,鱼肉因肌纤维较短,且富含水分,在贮藏过程中易因微生物滋生导致腐败变质,造成巨大损失。研究表明,希瓦氏菌属、假单胞菌属、气单胞菌属、弧菌属、发光杆菌属、芽孢杆菌属和部分肠杆菌等是海水鱼中的特定腐败菌(SSOs),在海水鱼的腐败过程中发挥着主导作用。同时,由部分食源性致病菌污染海水鱼,尤其是单增李斯特菌、沙门氏菌、溶血性大肠杆菌、副溶血性弧菌等对人体健康带来严重威胁。
乳酸菌是GRAS(Generally Regarded As Safety)级微生物,已广泛应用于食品发酵与保鲜。大量研究表明,部分乳酸菌如植物乳杆菌、嗜酸乳杆菌、干酪乳杆菌、棒状乳杆菌、德氏乳杆菌等可产生胞外细菌素(抗菌肽),这些不同来源的细菌素对革兰氏阳性菌、阴性菌以及真菌展示出不同的抑菌活性。乳酸菌源细菌素因抗菌活性强、安全性高等特点,是一类具有广阔应用前景的绿色生物防腐剂。目前,国内外关于Plantaricin、Lactococcin、Acidocin、Lactocin等乳酸菌源细菌素的分离纯化、结构鉴定、抑菌活性的研究已有大量报道,抗菌肽在线数据库中(The antimicrobial peptide database,2020年3月)已收录乳酸菌源细菌素43条。但是,大部分细菌素对革兰氏阳性菌(G+菌)有效,而对革兰氏阴性菌(G–菌)抗菌作用有限,只有植物乳杆菌素等少数细菌素可抑制G–菌。如植物乳杆菌所产的细菌素Plantaricin ST28MS和ST26MS可抑制铜绿假单胞菌和大肠杆菌;Plantaricin MG可以抑制鼠伤寒沙门氏菌和大肠杆菌;Plantaricin BacTN635可以抑制肠炎沙门氏菌,铜绿假单胞菌,哈夫尼菌属和沙雷氏菌属。至今,只有乳酸链球菌素(Nisin)和Pediocin PA-1两种乳酸菌源细菌素被批准可用作食品防腐剂,但在我国仅Nisin被允许可应用于食品。Nisin只对G+菌具有抑菌活性,对G–菌则无抑菌效果,限制了其在以腐败希瓦氏菌属、假单胞菌属等G–菌为特定腐败菌的生鲜海水鱼防腐保鲜中的应用。当前,国家对食品安全高度重视,因化学合成类防腐剂存在食品安全风险,开发新型、绿色、安全、广谱的乳酸菌源细菌素,用于海水鱼防腐保鲜,抑制微生物滋生与食源性致病菌污染具有重要意义。乳酸菌源广谱抗菌肽因抗菌活性强、抑菌谱广、安全性高等特点,是一类具有广阔应用前景的绿色生物防腐剂。
发明内容
本发明的目的在于针对现有技术的不足,提供一株产广谱抗菌肽的棒状乳杆菌及其抗菌肽的应用。
为实现上述目的,本发明采用如下技术方案:
一种产广谱抗菌肽的棒状乳杆菌,所述产广谱抗菌肽棒状乳杆菌为棒状乳杆菌(Lactobacillus coryniformis)FZU63,其分类命名为:棒状乳杆菌(Lactobacillus coryniformis),于2020年4月7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,保藏编号为:CGMCC NO. 19553,保藏地址为:北京市朝阳区北辰西路1号院3号。
上述棒状乳杆菌(Lactobacillus coryniformis)FZU63是从吉林省榆树市农村家庭自制酸白菜中分离获得的。
棒状乳杆菌(Lactobacillus coryniformis)FZU63革兰氏染色为阳性,菌落呈圆形,微白色,小而凸起,边缘整齐。
棒状乳杆菌(Lactobacillus coryniformis)FZU63产生的抗菌物质具有广谱的抑菌活性,对金黄色葡萄球菌ATCC 6538、金黄色葡萄球菌ATCC 12600、耐甲氧西林金黄色葡萄球菌MW2、耐甲氧西林金黄色葡萄球菌N315、单增李斯特菌CICC 21633、藤黄微球菌CMCC28001,6株革兰氏阳性菌和大肠埃希氏菌ATCC 25922、大肠埃希氏菌CMCC(B) 44102、肠道出血性大肠埃希氏菌CICC 21530、副溶血性弧菌ATCC 17802、猪霍乱沙门氏菌CICC 13312、鼠伤寒沙门氏菌CICC 51005、肠炎沙门氏菌CICC 21527、甲型副伤寒沙门氏菌CMCC(B)50093、乙型副伤寒沙门氏菌CMCC(B) 50094、阪崎克罗诺肠杆菌ATCC 29544、荧光假单胞菌AS1.55、希瓦氏菌 ATCC49138,12株革兰氏阴性菌,均表现出良好的抑菌效果。
上述产广谱抗菌肽棒状乳杆菌,即棒状乳杆菌(Lactobacillus coryniformis)FZU63产生的抗菌肽AMP-fzu63,所述抗菌肽AMP-fzu63包含2条多肽序列,具体多肽序列为:RQQPMTLDYRW、LVKNAKERGY。
上述抗菌肽AMP-fzu63在海水鱼保鲜中的应用。
本发明的优点在于:本发明筛选获得一株产广谱抗菌肽的棒状乳杆菌FZU63,棒状乳杆菌FZU63产生的抗菌肽对多种革兰氏阳性菌和阴性菌具有显著的抑制作用。棒状乳杆菌FZU63产生的抗菌肽AMP-fzu63包含多肽序列为RQQPMTLDYRW、LVKNAKERGY的2条多肽序列。棒状乳杆菌FZU63产生的抗菌肽AMP-fzu63应用于海水鱼保鲜中,能通过抑制海水鱼中微生物的生长与繁殖,从而降低不良气味物质己醛和1-辛烯-3-醇的产生,其作用效果显著优于现有技术中用于食品防腐保鲜的乳酸菌源抗菌肽乳酸链球菌素(Nisin)。
附图说明:
图1 PCR扩增产物凝胶电泳图。从左到右依次是FZU63、FZU72、FZU73、FZU101、FZU105、FZU122、Marker。
图2基于16S rDNA基因序列构建FZU63菌株的系统发育树。
图3 棒状乳杆菌FZU63发酵液对革兰式阳性菌的抑菌效果。A:金黄色葡萄球菌ATCC 6538, B:金黄色葡萄球菌ATCC 12600,C:耐甲氧西林金黄色葡萄球菌MW2,D:耐甲氧西林金黄色葡萄球菌N315,E:单增李斯特菌CICC 21633,F:藤黄微球菌CMCC 28001。
图4 棒状乳杆菌FZU63发酵液对革兰式阴性菌的抑菌效果。A:大肠埃希氏菌ATCC25922,B:大肠埃希氏菌CMCC(B) 44102,C:肠道出血性大肠埃希氏菌CICC 21530,D:副溶血性弧菌ATCC 17802, E:猪霍乱沙门氏菌CICC 13312,F:鼠伤寒沙门氏菌CICC 51005,G:肠炎沙门氏菌CICC 21527,H:甲型副伤寒沙门氏菌CMCC(B) 50093,I:乙型副伤寒沙门氏菌CMCC(B) 50094,J:阪崎克罗诺肠杆菌ATCC 29544,K:荧光假单胞菌AS1.55,L:希瓦氏菌ATCC49138。
图5棒状乳杆菌FZU63发酵液对真菌绿色木霉菌CGMCC3.2942的抑制效果。
图6样品的Sephadex LH-20凝胶层析图谱。
图7棒状乳杆菌FZU63发酵液粗提物反相HPLC层析图谱及不同分离组分的抑菌活性。
图8棒状乳杆菌FZU63发酵液粗提物反相HPLC层析不同分离组分的抑菌活性。图中C:流动相(20%甲醇);1、2、3、4、5为图7中洗脱峰1、2、3、4、5的收集产物。
图9抗菌肽AMP-fzu63中氨基酸序列为RQQPMTLDYRW多肽的二级质谱图。
图10抗菌肽AMP-fzu63中氨基酸序列为LVKNAKERGY多肽的二级质谱图。
图11抗菌肽AMP-fzu63在海鲈鱼冷藏保鲜中的应用效果。A,AMP-fzu63对海鲈鱼在冷藏保鲜过程中菌落总数的影响;B,AMP-fzu63对海鲈鱼在冷藏保鲜过程中鱼肉中己醛含量的影响;C,AMP-fzu63对海鲈鱼在冷藏保鲜过程中鱼肉中1-辛烯-3-醇含量的影响。
具体实施方式
为了使本发明所述的内容更加便于理解,下面结合具体实施方式对本发明所述的技术方案做进一步的说明,但是下述的实例仅仅是本发明其中的例子而已,并不代表本发明所限定的权利保护范围,本发明的权利保护范围以权利要求书为准。
实施例1
本发明中的棒状乳杆菌FZU63是从吉林省榆树市农村家庭自制酸白菜中分离获得的。
具体分离方法为:取10 g酸白菜经无菌破碎,使用无菌生理盐水重悬,10倍梯度稀释后,分别涂布于MRS固体培养基平板,37 ℃条件下静置培养18 h;挑取单菌落置于MRS液态培养基内,37 ℃条件下静置培养24 h,离心取上清,在排除有机酸、过氧化氢等物质干扰的情况下,分别以金黄色葡萄球菌和鼠伤寒沙门氏菌为指示菌通过固体琼脂打孔抑菌法,筛选出产抑菌物质的菌株。
筛选所得产抑菌物质的菌株革兰氏染色为阳性,菌落呈圆形,微白色,小而凸起,边缘整齐,这些特点与典型乳酸菌菌落特征相同。从中挑选出抑菌圈直径在10 mm以上的6株菌将其分别命名为FZU63、FZU72、FZU73、FZU101、FZU105、FZU122,通过GB4789.35-2016鉴定这6株菌,鉴定结果见表1,初步判定为乳酸菌。
表1 初筛6株菌株生化法反应检测结果
注:+表示菌株呈阳性。
FZU63菌株接种于MRS液体培养基,37℃条件下经活化、转接培养至对数生长期。取1~2 mL培养液10000 g/min离心1 min,弃上清,菌体细胞使用商业细菌总DNA提取试剂盒,参照说明书提取总DNA。采用16S rDNA基因引物 16SAF (5'-AGAGTTTGATCCTGGCTCAG-3′)和16SAR (5′-TACGGYTACCTTGTTACGACTT-3′)进行PCR扩增,取5μL PCR产物进行1%琼脂糖凝胶电泳,电泳结果见图1。
用试剂盒纯化回收PCR产物,由生工生物工程(上海)股份有限公司进行16S rRNA全序列测序。菌株FZU63的16S rDNA序列共1242bp,测序结果如SEQ ID NO.1所示。
SEQIDNO.1:
CTTTGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGTTAGTTGGTAAGGTAACGGCTTACCAAGACAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTAGGCTCGTAAAACTCTGTTGTTAGAGAAGAACACATATGAGAGTAACTGTTCATCTTTTGACGGTATTCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTTAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTTTGCCCATCGTAGAGATAAAACTTTCCCTTCGGGGACAAAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTACCAGTTGCCAGCATTTAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGTTGCGAACTCGCGAGGGTAAGCTAATCTCTTAAAGCCGATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCT
将FZU63菌株的16S rDNA序列在NCBI中(https://www.ncbi.nlm.nih.gov/)使用BLAST比对。综合生理生化鉴定以及16S rDNA同源性比较结果可以把该菌株归类为棒状乳杆菌(Lactobacillus coryniformis),构建系统发育树见图2。
实施例2
棒状乳酸菌FZU63接种于MRS液态培养基内,37 ℃条件下静置培养24 h,离心取上清,分别以金黄色葡萄球菌ATCC 6538、金黄色葡萄球菌ATCC 12600、耐甲氧西林金黄色葡萄球菌MW2、耐甲氧西林金黄色葡萄球菌N315、单增李斯特菌CICC 21633、藤黄微球菌CMCC28001、大肠埃希氏菌ATCC 25922、大肠埃希氏菌CMCC(B) 44102、肠道出血性大肠埃希氏菌CICC 21530、副溶血性弧菌ATCC 17802、猪霍乱沙门氏菌CICC 13312、鼠伤寒沙门氏菌CICC51005、肠炎沙门氏菌CICC 21527、甲型副伤寒沙门氏菌CMCC(B) 50093、乙型副伤寒沙门氏菌CMCC(B) 50094、阪崎克罗诺肠杆菌ATCC 29544、荧光假单胞菌AS1.55、希瓦氏菌ATCC49138、绿色木霉菌CGMCC3.2942为指示菌,细菌接种于LB固体琼脂,绿色木霉菌接种于PDA固体培养基,采用打孔抑菌法评价抗菌剂对指示菌的抑菌效果。如表2所示,琼脂孔扩散法实验结果表明,棒状乳杆菌FZU63发酵上清液具有较为广谱的抑菌活性,对18株细菌指示菌均表现出良好的抑菌效果,包括6株革兰氏阳性菌(见图3)和12株革兰氏阴性菌(见图4),对真菌没有抑菌效果(见图5)。
表2 棒状乳杆菌FZU63发酵液对不同指示菌抑菌抑菌圈直径
实施例3
棒状乳杆菌FZU63经MRS液体培养基,37 ℃条件下静态厌氧培养36 h,离心取上清,加入3倍体积乙醇,4 ℃静置12 h后离心取上清,并通过真空冷凝旋转蒸发仪浓缩并脱除乙醇。将经过浓缩的样品取5 mL加入到Sephadex LH-20凝胶柱,用20%甲醇等度洗脱,流速设置为0.2 mL/min,每管收集2 mL,验证各管是否具有抑菌活性及活性大小,洗脱曲线如图6。将具有抑菌活性的分管收集并采用旋转蒸发仪浓缩,置于4 ℃保存备用。
由Sephadex LH-20过滤层析获得棒状乳杆菌FZU63发酵上清液粗提物,图7为该粗提物的反相HPLC层析图谱,收集不同保留时间的洗脱峰(1、2、3、4、5),用旋转蒸发仪浓缩,以鼠伤寒沙门氏菌CICC21483为指示菌测其抑菌活性得图8,峰1、3和4均具有抑菌活性。保留时间为9.193 min的活性峰4抑菌活性最强,其抑菌圈直径达18.82 mm,用微量核酸蛋白测定仪测其蛋白质含量为4.426 mg/mL。收集该活性峰经高分辨质谱进行下一步结构鉴定。
实施例4
经高效液相色谱系统(HPLC)分离纯化获得的具有抑菌活性的分离组分进一步经纳升四级杆飞行时间串联质谱(NLM)分析对抗菌肽AMP-fuz63的分子结构进行鉴定。如表3示所示,经质谱鉴定获得2条多肽序列,经在线抗菌肽数据库(Antimicrobial peptidedatabase: http://aps.unmc.edu/AP/main.php)比对,未发现相同序列。表明,鉴定出2条全新的棒状乳杆菌源抗菌肽(2条抗菌肽的二级质谱图谱见图9、图10)。
表3 抗菌肽AMP-fuz63经质谱鉴定后的氨基酸序列
实施例5
鲜活海鲈鱼经处死、去除鱼鳞、内脏并分割称重后置于无菌密封袋,根据鱼肉重量以128 μg/g加入抗菌肽AMP-fzu63,混合均匀后密封,置于4 ℃条件下储藏,同时设置无菌生理盐水处理与Nisin处理对照组,并分别于3、7、11、15天取样。
(1)将鱼肉与生理盐水以1:10混合,均质后经梯度稀释,涂布于LB固体培养基,置于37 ℃条件下培养18 h后进行菌落计数。
(2)称重3~4 g鱼肉,剪碎后置于12 mL顶空萃取瓶中,加入5 mL超纯水、1 g NaCl、磁力转子和10 μL 10 mg/L 2-辛醇(内标物),用密封垫迅速密封样品瓶,置于磁力搅拌加热台上,搅拌速度600 r/min,在40 ℃水浴下孵化20 min,然后插入老化后的固相微萃取针,顶出萃取纤维,使其固定在距离液面0.5~1.0 cm处,边搅拌边顶空吸附,萃取30 min后收回萃取纤维,拔出萃取针,插入气相色谱质谱联用仪中分析。第一次使用萃取头萃取前,要在250 ℃将萃取头老化至无杂峰再进行萃取,并经气相色谱法质谱联用仪测定鱼肉中挥发性气味物质。气相色谱条件:进样口温度250 ℃;柱温箱起始温度40 ℃,保留8 min,然后以4 ℃/min升温至150 ℃,再以20 ℃/min升温至250 ℃,保留5 min;不分流进样,载气:99.999%高纯度氦气,载气流速:1 mL/min。质谱条件:离子源温度为 230 ℃,电离方式为EI,电离能量为 70 eV,接口温度为 250 ℃,四级杆温度150 ℃,选择SCAN模式为扫描方式进行定性分析,离子碎片的扫描范围为 30~500 m/z,溶剂延迟时间为2.5 min。
由图11A所示,随冷藏时间延长,抗菌肽AMP-fzu63处理组海鲈鱼鱼肉中的菌落总数显著低于生理盐水与Nisin处理组。表明AMP-fzu63可有效抑制鱼肉中微生物的滋生,延缓海鲈鱼因微生物生长繁殖导致的腐败变质。
由图11B和11C所示,海鲈鱼储藏期间鱼肉中不良气味物质己醛和1-辛烯-3-醇含量呈现出随冷藏时间延长先递增后下降的变化趋势,且在7-9日时达到最大值。在抗菌肽AMP-fzu63处理组,鱼肉中的己醛和1-辛烯-3-醇含量随冷藏时间延长显著低于生理盐水与Nisin处理组。有研究表明鱼肉中己醛和1-辛烯-3-醇含量的变化与微生物的生长活动具有密切关联。由此可见,抗菌肽AMP-fzu63能通过抑制海鲈鱼鱼肉中微生物的生长与繁殖,从而降低不良气味物质己醛和1-辛烯-3-醇的产生。
以上所述仅为本发明的较佳实施例,凡依本发明申请专利范围所做的均等变化与修饰,皆应属本发明的涵盖范围。
SEQUENCE LISTING
<110> 福州大学
<120> 一株产广谱抗菌肽的棒状乳杆菌及其抗菌肽的应用
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acggcccaaa ctcctacggg aggcagcagt agggaatctt ccacaatgga cgaaagtctg 180
atggagcaac gccgcgtgag tgaagaaggg tttaggctcg taaaactctg ttgttagaga 240
agaacacata tgagagtaac tgttcatctt ttgacggtat tcaaccagaa agccacggct 300
aactacgtgc cagcagccgc ggtaatacgt aggtggcaag cgttgtccgg atttattggg 360
cgtaaagcga gcgcaggcgg ttttttaagt ctgatgtgaa agccttcggc ttaaccgaag 420
aagtgcatcg gaaactggga aacttgagtg cagaagagga cagtggaact ccatgtgtag 480
cggtgaaatg cgtagatata tggaagaaca ccagtggcga aggcggctgt ctggtctgta 540
actgacgctg aggctcgaaa gcatgggtag cgaacaggat tagataccct ggtagtccat 600
accgtaaacg atgaatgcta agtgttggag ggtttccgcc cttcagtgct gcagctaacg 660
cattaagcat tccgcctggg gagtacgacc gcaaggttga aactcaaagg aattgacggg 720
ggcccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag 780
gtcttgacat cctttgccca tcgtagagat aaaactttcc cttcggggac aaagtgacag 840
gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 900
gcaaccctta ttaccagttg ccagcattta gttgggcact ctggtgagac tgccggtgac 960
aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac 1020
acgtgctaca atggacggta caacgagttg cgaactcgcg agggtaagct aatctcttaa 1080
agccgatctc agttcggatt gtaggctgca actcgcctac atgaagtcgg aatcgctagt 1140
aatcgcggat cagcacgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1200
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Claims (3)
1.一株产广谱抗菌肽的棒状乳杆菌,其特征在于:所述产广谱抗菌肽的棒状乳杆菌为棒状乳杆菌(Lactobacillus coryniformis)FZU63,其分类命名为:棒状乳杆菌(Lactobacillus coryniformis),于2020年4月7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,保藏编号为:CGMCC NO. 19553,保藏地址为:北京市朝阳区北辰西路1号院3号。
2.如权利要求1所述的一株产广谱抗菌肽的棒状乳杆菌产生的抗菌肽组合物AMP-fzu63,其特征在于:所述抗菌肽组合物AMP-fzu63包含2条多肽,所述2条多肽的氨基酸序列分别为: RQQPMTLDYRW、LVKNAKERGY。
3.如权利要求2所述的抗菌肽组合物AMP-fzu63在海水鱼保鲜中的应用。
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