食品科学 ›› 2022, Vol. 43 ›› Issue (2): 134-141.doi: 10.7506/spkx1002-6630-20201029-294

• 生物工程 • 上一篇    下一篇

海洋细菌Pseudoalteromonas sp. HL9的筛选鉴定及产磷脂酶B性质

王藏,马小艺,田小鹏,祖航天,丁延帅,吕明生,王淑军   

  1. (1.江苏海洋大学 海洋资源与环境重点实验室,海洋生物技术重点实验室,江苏 连云港 222005;2.江苏海洋大学 江苏省海洋生物产业技术协同创新中心,江苏 连云港 222005)
  • 出版日期:2022-01-25 发布日期:2022-01-29
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2018YFC0311106);江苏省高等教育优势学科项目(PAPD); 江苏省研究生创新项目(KYCX19-2297)

Screening and Identification of Marine Bacterium Pseudoalteromonas sp. HL9 and Characteristics of Phospholipase B Produced by It

WANG Cang, MA Xiaoyi, TIAN Xiaopeng, ZU Hangtian, DING Yanshuai, LÜ Mingsheng, WANG Shujun   

  1. (1. Jiangsu Key Laboratory of Marine Bioresources and Environment, Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, China; 2. Co-innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang 222005, China)
  • Online:2022-01-25 Published:2022-01-29

摘要: 从黄海海泥中筛选产磷脂酶菌株,其中酶活力最高的HL9鉴定为假交替单胞菌属(Pseudoalteromonas)。该菌株最佳生长培养基组成为米糠与酵母粉,最佳生长pH 8.0、NaCl质量浓度30 g/L、培养温度25 ℃。优化后产酶最高发酵条件:发酵培养基配方为10 g/L麸皮和5 g/L鱼粉蛋白胨,NaCl 20 g/L,pH 8.5,接种量2%,30 ℃培养48 h。薄层层析和气相色谱分析表明,菌株HL9产磷脂酶B,且磷脂酶B水解sn-2酰基的产物占70%。该酶的最适反应温度和pH值分别为45 ℃和7.5。Ca2+、Co2+、Mn2+等金属离子对酶活力有促进作用,Cu2+对酶活力有抑制作用。磷脂酶B在有机溶剂中可以保持80%的酶活力。反应温度高和碱性条件等催化特性使该酶具有较好应用前景,研究结果也为进一步探索该酶的结构与功能奠定基础。

关键词: 假交替单胞菌;磷脂酶B;酶学性质

Abstract: In this study, we screened and identified Pseudoalteromonas sp. HL9 with the capacity to produce phospholipase B from the mud of the Yellow Sea. The optimal growth medium for this strain consisted of rice bran, yeast powder, and 30 g/L NaCl at pH 8.0, and the optimal culture temperature was 25 ℃. The optimal fermentation conditions that provided maximum phospholipase B activity were found to be culture at 30 ℃ for 48 h in a medium containing rice bran 10 g/L, fish meal peptone 5 g/L, and NaCl 20 g/L at pH 8.5. Thin layer chromatography (TLC) and gas chromatography (GC) results showed that strain HL9 could produce phospholipase B, and that the hydrolysate of sn-2 acyl groups with the enzyme accounted for 70% of the total. The optimal temperature and pH of this enzyme were 45 ℃ and 7.5, respectively. Ca2+, Co2+ and Mn2+ could enhance the activity of phospholipase B, while Cu2+ had an inhibitory effect on the enzyme activity. The phospholipase B retained 80% of its activity in organic solvents. In summary, this enzyme has good catalytic properties at high temperature or alkaline conditions and thus has broad application prospects. The results of this study will lay the foundation for investigating its structure and function.

Key words: Pseudoalteromonas; phospholipase B; characteristics

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