摘要
为有效利用烟草废弃物生产烟用酶制剂,以烟草源耐烟碱木质素降解菌枯草芽孢杆菌(Bacillussubtilis)YY-10为研究对象,通过遗传和分泌蛋白组成分析、基因编辑,研究其作为产酶宿主菌的可行性。结果表明:①枯草芽孢杆菌YY-10基因组含有1个环形染色体,编码4252个基因,含多种碳水化合物活性酶,分泌蛋白中富含果胶裂解酶和淀粉酶等降解酶。②以YY-10为宿主菌构建的产果胶酶基因工程菌(YY-10-pelA)可实现目标酶蛋白的表达与分泌,酶活性较野生菌提高约3倍,且具有在营养胁迫条件下利用碱性木素生长的能力。③对YY-10菌株与传统枯草芽孢杆菌宿主菌产酶能力及酶制剂的应用效果的分析发现,YY-10菌株作为烟用酶制剂产酶宿主菌在提高烟草废弃物利用率和烟草的感官舒适性方面具有明显优势。
In order to promote effective utilization of tobacco waste and sustainable production of tobaccoenzyme additives, Bacillus subtilis YY-10, a nicotine-resistant lignin-degrading bacterium derived fromtobacco, was investigated by genome sequencing, exoprotein composition analysis and gene editingtechniques to evaluate its feasibility as an enzyme-producing host. The results showed that: 1)The YY-10genome had a circular chromosome encoding 4 252 genes, which contained various active carbohydrateenzymes. The exoproteins of YY-10 were rich in pectate lyase and amylase. 2)A pectate lyase expressionsystem was developed and introduced into the YY-10 host, resulting in a genetically engineered strain,YY-10-pelA. The strain achieved expression and secretion of the target enzyme, and exhibited enzymeactivity about three times higher than the wild type, and demonstrated the ability to utilize alkaline ligninfor growth under nutrient stress. 3) Compared with well-established B. subtilis hosts, YY-10 as an enzyme-producing host using tobacco waste for enzyme preparation showed considerable enzymeexpression capacity and outstanding advantages in improving the sensory quality of tobacco.
作者
程成
陈浩锋
陈善义
李菁菁
何伟
龙腾
陈义强
邓小华
周康熙
姜振锟
丁宁
袁吉锋
范坚强
CHENG Cheng;CHEN Haofeng;CHEN Shanyi;LI Jingjing;HE Wei;LONG Teng;CHEN Yiqiang;DENG Xiaohua;ZHOU Kangxi;JIANG Zhenkun;DING Ning;YUAN Jifeng;FAN Jianqiang(Technology Center,China Tobacco Fujian Industrial Co.,Ltd.,Xiamen 361021,Fujian,China;School of Life Sciences,Xiamen University,Xiamen 361102,Fujian,China)
出处
《烟草科技》
北大核心
2025年第7期20-27,共8页
Tobacco Science & Technology
基金
福建中烟工业有限责任公司博士后项目“烟草源枯草芽孢杆菌的定向改造与优化”(FJZYKJJH2023BH003)。
