The Cloning and Bioinformatics Analysis of Mannanase Gene in Lactobacillus casei HDS-01

doi:10.11924/j.issn.1000-6850.casb2023-0165

Abstract

Abstract:

The purpose of this study was to explore the structure and function of mannanase gene by gene cloning and bioinformatics. The mannanase gene M1 was obtained from genome of Lactobacillus casei HDS-01 through local BLAST. The differential expression of M1 in MRS medium and KGM medium was determined by qRT-PCR, indicating that Konjac powder in KGM medium was better in inducing M1 expression. Gene M1 was cloned from genomic DNA of L. casei HDS-01, and its sequence was predicted and analyzed by bioinformatics method. The sequence was 2640 bp and could be translated into 879 amino acids, including a 2640 bp open reading frame. The molecular weight of M1 protein was 98723.55 Da and the isoelectric point was 5.58. The secondary structure of the stable M1 protein consisted of α-helix, β-folding, β-turning and random curling. The results not only provide theoretical basis for the molecular modification of mannanase, but also help to understand the metabolic pathway of mannan in lactic acid bacteria.

Key words: Lactobacillus casei, bioinformatics analysis, clone, mannanase, protein secondary structure

NING Yingying, ZHANG Xin, ZHAO Dan. The Cloning and Bioinformatics Analysis of Mannanase Gene in Lactobacillus casei HDS-01[J]. Chinese Agricultural Science Bulletin, 2023, 39(26): 33-40.

Figures/Tables 11

References 26

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时间/h	组别	CT值		△CT	△△CT	2^-△△^CT
时间/h	组别	CT-M1	CT-16S	△CT	△△CT	2^-△△^CT
12	MRS	26.07±0.19	26.06±0.25	0.01	0	1.00^a
12	KGM	27.70±0.22	25.46±0.35	2.24	2.23	0.21^b
24	MRS	27.58±0.27	25.15±0.10	2.43	2.42	0.19^b
24	KGM	24.98±0.31	23.31±0.28	1.67	1.67	0.32^b
36	MRS	27.62±0.11	26.69±0.32	0.94	0.93	0.53^b
36	KGM	28.46±0.23	28.85±0.24	-0.39	-0.39	2.49^b
48	MRS	27.75±0.16	27.08±0.14	0.67	0.67	0.63^b
48	KGM	28.58±0.30	27.94±0.26	0.64	0.63	1.63^b
60	MRS	28.52±0.26	28.43±0.21	0.09	0.08	1.03^a
60	KGM	28.17±0.22	28.20±0.33	-0.04	-0.04	1.30^b
72	MRS	28.24±0.18	28.93±0.17	-0.69	-0.70	0.64^b
72	KGM	31.87±0.13	33.19±0.15	-1.33	-1.33	0.95^b

时间/h	组别	CT值		△CT	△△CT	2^-△△^CT
时间/h	组别	CT-M1	CT-16S	△CT	△△CT	2^-△△^CT
12	MRS	26.07±0.19	26.06±0.25	0.01	0	1.00^a
12	KGM	27.70±0.22	25.46±0.35	2.24	2.23	0.21^b
24	MRS	27.58±0.27	25.15±0.10	2.43	2.42	0.19^b
24	KGM	24.98±0.31	23.31±0.28	1.67	1.67	0.32^b
36	MRS	27.62±0.11	26.69±0.32	0.94	0.93	0.53^b
36	KGM	28.46±0.23	28.85±0.24	-0.39	-0.39	2.49^b
48	MRS	27.75±0.16	27.08±0.14	0.67	0.67	0.63^b
48	KGM	28.58±0.30	27.94±0.26	0.64	0.63	1.63^b
60	MRS	28.52±0.26	28.43±0.21	0.09	0.08	1.03^a
60	KGM	28.17±0.22	28.20±0.33	-0.04	-0.04	1.30^b
72	MRS	28.24±0.18	28.93±0.17	-0.69	-0.70	0.64^b
72	KGM	31.87±0.13	33.19±0.15	-1.33	-1.33	0.95^b