Cloning, characterization, and antifungal activity of an endo-1,3-beta-D: -glucanase from Streptomyces sp. S27.
An endo-1,3-beta-D: -glucanase gene, designated as bglS27, was cloned from Streptomyces sp. S27 and successfully expressed in Escherichia coli BL21 (DE3). The full-length gene contains 1,362 bp and encodes a protein of 453 amino acids with a calculated molecular mass of 42.7 kDa. The encoded protein comprises a catalytic module of glycosyl hydrolase family 16, a short glycine linker region, and a family 13 carbohydrate-binding module. The purified recombinant enzyme (BglS27) showed optimal activity at 65 degrees C and pH 5.5 and preferentially catalyzed the hydrolysis of glucans with a beta-1,3-linkage using an endolytic mode of action. The specific activity and K(m) value of BglS27 for laminarin were 236.0 U mg(-1) and 1.89 mg ml(-1), respectively. In antifungal assay, BglS27 had the ability to inhibit the growth of phytopathogenic fungi Rhizoctonic solani and Fusarium oxysporum and some mycotoxin-producing fungi Fusarium crookwellense and Paecilomyces variotii. These favorable properties make BglS27 a good candidate for utilization in biotechnological applications such as plant protection, feed, and food preservation.
Shi
Pengjun
P
Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
Yao
Guoyu
G
Yang
Peilong
P
Li
Ning
N
Luo
Huiying
H
Bai
Yingguo
Y
Wang
Yaru
Y
Yao
Bin
B
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005658:Fungi / Q000187:drug effects*; D005798:Genes, Bacterial; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D005936:Glucans / Q000378:metabolism; D006868:Hydrolysis; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D016415:Sequence Alignment; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D000935:Antifungal Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005936:Glucans; D011134:Polysaccharides; D011994:Recombinant Proteins; C008247:laminaran; D004693:Glucan Endo-1,3-beta-D-Glucosidase