Cloning and sequencing of the genes encoding cyclic tetrasaccharide-synthesizing enzymes from Bacillus globisporus C11

• Published in: Bioscience, biotechnology, and biochemistry Vol. 66; no. 5; pp. 1057 - 1068
• Main Authors: Aga, H. (Hayashibara Biochemical Labs. Inc., Okayama (Japan)), Maruta, K, Yamamoto, T, Kubota, M, Fukuda, S, Kurimoto, M, Tsujisaka, Y
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.05.2002; Japan Society for Bioscience Biotechnology and Agrochemistry
• Subjects: 6-glucosyltransferase; Amino Acid Sequence; AMINO ACID SEQUENCES; BACILLUS; Bacillus - enzymology; Bacillus globisporus; Base Sequence; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Cloning, Molecular; cyclic tetrasaccharide; DNA, Bacterial; Fundamental and applied biological sciences. Psychology; GENES; Genetics; GLUCANS; GLUCOSIDASES; Glucosyltransferases - chemistry; Glucosyltransferases - genetics; isomaltosyltransferase; Mission oriented research; MOLECULAR CLONING; Molecular Sequence Data; NUCLEOTIDE SEQUENCE; Oligosaccharides - biosynthesis; Open Reading Frames; Polymerase Chain Reaction; POLYSACCHARIDES; Sequence Homology, Amino Acid; TRANSFERASES; Bacillales; Gene; Nucleotide sequence; Enzyme; Cyclic compound; Transferases; Bacteria; Homology; Bacillaceae; Structure function relationship; Tetrasaccharide
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.66.1057

The genes for isomaltosyltransferase (CtsY) and 6-glucosyltransferase (CtsZ), involved in synthesis of a cyclic tetrasaccharide from alpha-glucan, have been cloned from the genome of Bacillus globisporus C11. The amino-acid sequence deduced from the ctsY gene is composed of 1093 residues having a signal sequence of 29 residues in its N-terminus. The ctsZ gene encodes a protein consisting of 1284 residues with a signal sequence of 35 residues. Both of the gene products show similarities to alpha-glucosidases belonging to glycoside hydrolase family 31 and conserve two aspartic acids corresponding to the putative catalytic residues of these enzymes. The two genes are linked together, forming ctsYZ. The DNA sequence of 16,515 bp analyzed in this study contains four open reading frames (ORFs) upstream of ctsYZ and one ORF downstream. The first six ORFs, including ctsYZ, form a gene cluster, ctsUVWXYZ. The amino-acid sequences deduced from ctsUV are similar in to a sequence permease and a sugar-binding protein for the sugar transport system from Thermococcus sp. B1001. The third ctsW encodes a protein similar to CtsY, suggested to be another isomaltosyltransferase preferring panose to high-molecular-mass substrates.