摘要
方斑东风螺是我国及东南亚地区重要的海水养殖品种,由于养殖底质污染,导致方斑东风螺病害频发。文章从健康方斑东风螺养殖池底沙中筛选蛋白酶高产菌株,基于形态、生理生化和分子特征对高产蛋白酶菌株进行鉴定,通过菌株溶血性检测和药敏试验初步评价其安全性,在单因素试验的基础上通过正交试验对其发酵培养基碳源、氮源及金属盐进行优化,并在室内养殖条件下考察了其对方斑东风螺养殖水体(氨氮、亚硝酸盐氮、化学需氧量)、底沙(总氮、有机碳)以及方斑东风螺生长(存活率、特定生长率)的影响。结果表明:从健康方斑东风螺养殖池底沙分离得到一株高产蛋白酶芽孢菌,其蛋白酶活性为324.0U·mL^(-1),命名为YZS02,经鉴定菌株YZS02为越南蔷薇菌(Rossellomorea vietnamensis),该菌株没有溶血活性,对24种抗生素敏感,其最优发酵培养基为酵母粉15g·L^(-1)、可溶性淀粉20g·L^(-1)、MgCl28g·L^(-1)、NaCl 30g·L^(-1),用此优化培养基在30℃,200r·min-1条件下摇瓶发酵24h,菌体密度可达到7.19×10^(9)个·毫升^(-1),该密度是优化前的2.89倍,同时优化后蛋白酶活性可达753.3U·mL^(-1),为原来的2.3倍。YZS02能够有效降低方斑东风螺养殖水体氨氮和化学需氧量水平,对底沙中的总氮也有很好的去除效果,试验结束时对照组水体氨氮浓度为2.33mg·L^(-1)、化学需氧量为278.37mg·L^(-1)、底沙总氮含量为538.31μg·g-1,而试验组水体氨氮浓度为0.78 mg·L^(-1)、化学需氧量为128.47mg·L^(-1)、底沙总氮含量为128.71μg·g-1。YZS02还能有效提高方斑东风螺的存活率及特定生长率。
Babylonia areolata is an important marine aquaculture variety in China and Southeast Asia.Due to the pollution of the breeding substrate,the disease of Babylonia areolata happens frequently.In this study,high-yielding protease strains were screened from the bottom sand of the healthy Babylonia areolata breeding pond.The high-yielding protease strain was identified based on morphology,physiological,biochemical and molecular characteristics.Its safety was preliminarily evaluated through strain hemolytic detection and drug sensitivity test,and its fermentation medium was optimized by single-factor and orthogonal tests.Chemicalization,and under indoor aquaculture conditions,the effects of the water bodies(ammonia nitrogen,nitrite nitrogen,COD),sediment(total nitrogen,organic carbon)and the growth(survivability rate and specific growth rate)of snails were investigated.The results showed that one strain YZS02 of high-yielding protease bacillus was isolated from the bottom sand of one healthy Babylonia areolata breeding pond,and was identified as Rossellomorea vietnamensis.The strain has no hemolysis activity.Sensitive to 24 antibiotics,the optimal fermentation medium is composed of 15 g·L^(-1) yeast powder,soluble starch 20 g·L^(-1),MgCl28 g·L^(-1),sodium chloride 30 g·L^(-1).This optimized liquid medium is used to shake bottle fermentation for 24 hours at 30℃and 200 r·min-1.Under this conditions,the bacterial density can reach 7.19×10^(9) cfu·mL^(-1),which is 2.89 times than that before optimization.YZS02 can effectively reduce the level of ammonia nitrogen and COD in the water cultured by Babylonia areolata.It also has a postive effect on removing the total nitrogen in the bottom sand,which can effectively improve the survival rate and specific growth rate of Babylonia areolata.At the end of the experiment,the ammonia nitrogen concentration in the control group was 2.33 mg·L^(-1),COD was 278.37 mg·L^(-1),and the total nitrogen content of sis was 538.31μg·g-1,while the ammonia nitrogen concentration in th
作者
武慧宇
高秀菊
葛东振
朱军
邹潇潇
黄惠琴
顾志峰
鲍时翔
WU Huiyu;GAO Xiuju;GE Dongzhen;ZHU Jun;ZOU Xiaoxiao;HUANG Huiqin;GU Zhifeng;BAO Shixiang(College of Oceanography,Hainan University,Haikou 570228,China;Institute of Tropical Bioscience and Biotechnology,CATAS&Key Laboratory of Biology and Genetic Resources of Tropical Crops of Hainan Province,Hainan Institute for Tropical Agricultural Resources,Haikou 571101,China;Zhanjiang Experimental Station,CATAS,Zhanjiang 524013,China;Hainan Provincial Key Laboratory for Functional Components Research and Utilization of Marine Bio-resources,CATAS,Haikou 571101,China)
出处
《热带海洋学报》
CAS
CSCD
北大核心
2023年第5期124-133,共10页
Journal of Tropical Oceanography
基金
农业农村部财政专项(NFZX2021、NHYYSWZZZYKZX2020)
中央级公益性科研院所基金项目(1630052019014)。
关键词
方斑东风螺
养殖
越南蔷薇菌
蛋白酶
培养基优化
Babylonia areolata
aquaculture
Rossellomorea vietnamensis
protease
Medium optimization
