Screening and Identification of Marine Electricity-producing and Lipase-producing Bacteria and Preliminary Study on Its Culture Conditions

doi:10.13560/j.cnki.biotech.bull.1985.2020-0833

Abstract

Abstract:

The objective of this work is to study the characteristics and electricity-producing of lipase-producing bacteria in the sea. The strains with lipase-producing feature were screened from the intertidal marine mud in Zhoushan sea area. Olive oil was used as the sole carbon source for plate screening,NaOH titration was used to determine the lipase activity,and the effect of fermentation condition on the lipase activity was preliminarily studied. To determine the taxonomic status of strains,16S rRNA gene analysis was used for identification,phylogenetic tree was constructed,and the effects of various culture conditions on the enzyme production activity were preliminarily studied through single-factor experiment. The microbial electric performance was preliminarily explored. The results showed that two strains with lipase-producing activity were selected from the intertidal mud,labeled S11 and S22,respectively. Molecular identification results revealed that S11 and Pseudomonas putidawa WAB1947,S22 and Bacillus substills LSRB were the closest in phylogenetic status respectively. The results of single-factor experiment demonstrated that the corresponding enzyme activity of the two strains was the best when the culture time was 48 h,the initial pH value was 7.0,and the urea content was 0.35 g/L. Microbial fuel cell（MFC）was constructed for voltage acquisition,and the results showed that the maximum output voltage of S11 and S22 was 0.055 V and 0.031 V,respectively. In conclusion,strain S11 and S22 is Pseudomonas putida and Bacillus substills,respectively,and they present certain electricity-producing capacity.

Key words: lipase, 16S rRNA, phylogenetic tree, microbial fuel cell

HUANG Yang-tian, LU Yu-biao, HUANG Yi-tie, MENG Fan-long, XU Kai-wen, LI Peng. Screening and Identification of Marine Electricity-producing and Lipase-producing Bacteria and Preliminary Study on Its Culture Conditions[J]. Biotechnology Bulletin, 2020, 36(12): 91-97.

Figures/Tables 10

References 28

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操作步骤	空白对照/mL	样品/mL（3个重复）
0.05 mol/L pH8.0 Tris-HCl缓冲溶液	5	5
橄榄油乳化液	4	4
	60℃水浴预热5-10 min
95%无水乙醇	15	0
发酵液	1.0（灭活）	1.0
	60℃水浴、震荡10 min
95%无水乙醇	0	15
1%酚酞	0.025	0.025
0.05 mol/L NaOH溶液滴定至终点	V_空	V_样

操作步骤	空白对照/mL	样品/mL（3个重复）
0.05 mol/L pH8.0 Tris-HCl缓冲溶液	5	5
橄榄油乳化液	4	4
	60℃水浴预热5-10 min
95%无水乙醇	15	0
发酵液	1.0（灭活）	1.0
	60℃水浴、震荡10 min
95%无水乙醇	0	15
1%酚酞	0.025	0.025
0.05 mol/L NaOH溶液滴定至终点	V_空	V_样