Cloning and expression of a Streptomyces plicatus chitinase (chitinase-63) in Escherichia coli - PubMed (original) (raw)
. 1988 Jan 5;263(1):443-7.
Affiliations
- PMID: 3275646
Free article
Cloning and expression of a Streptomyces plicatus chitinase (chitinase-63) in Escherichia coli
P W Robbins et al. J Biol Chem. 1988.
Free article
Abstract
4-Methylumbelliferyl (4-MU) glycosides of N-acetylglucosamine oligosaccharides were used as substrates to detect expression of a Streptomyces chitinase in Escherichia coli. Low levels of enzyme were detected when S. plicatus DNA was cloned into a bacteriophage lambda vector (EMBL-4). Subcloning into E. coli plasmids also gave low but detectable levels of enzyme expression. High level expression was achieved by resection of the cloned S. plicatus DNA with Bal31 followed by in-frame fusion to the amino-terminal peptide sequence of beta-galactosidase found in the pUC vectors. The Streptomyces chitinase was secreted into the periplasmic space of E. coli, and its signal sequence was removed. We characterized the activity of the cloned enzyme and compared it to three other purified Streptomyces plicatus chitinases with respect to hydrolysis of the 4-MU oligosaccharides. We found that two of the enzymes form 4-methylumbelliferone much more rapidly from the 4-MU disaccharide than from the trisaccharide. These same enzymes convert the 4-MU trisaccharide primarily to diacetylchitobiose and the 4-MU monosaccharide, a nonfluorescent product. The latter compound is not hydrolyzed appreciably by any of the enzymes. On the basis of these results, we suggest a new definition of "exo" and "endo" chitinase that differs from that found in the literature. We propose that exochitinase activity be defined as processive action starting at the nonreducing ends of chitin chains with release of successive diacetylchitobiose units, and that endochitinase activity be defined as random cleavage at internal points in chitin chains.
Similar articles
- Cloning and high-level expression of chitinase-encoding gene of Streptomyces plicatus.
Robbins PW, Overbye K, Albright C, Benfield B, Pero J. Robbins PW, et al. Gene. 1992 Feb 1;111(1):69-76. doi: 10.1016/0378-1119(92)90604-n. Gene. 1992. PMID: 1532161 - Molecular cloning and expression of the gene encoding family 19 chitinase from Streptomyces sp. J-13-3.
Okazaki K, Yamashita Y, Noda M, Sueyoshi N, Kameshita I, Hayakawa S. Okazaki K, et al. Biosci Biotechnol Biochem. 2004 Feb;68(2):341-51. doi: 10.1271/bbb.68.341. Biosci Biotechnol Biochem. 2004. PMID: 14981297 - Cloning and sequence analysis of the gene encoding a thermostable chitinase from Streptomyces thermoviolaceus OPC-520.
Tsujibo H, Endo H, Minoura K, Miyamoto K, Inamori Y. Tsujibo H, et al. Gene. 1993 Nov 30;134(1):113-7. doi: 10.1016/0378-1119(93)90183-4. Gene. 1993. PMID: 8244021 - Expression in Escherichia coli of a gene encoding a thermostable chitinase from Streptomyces thermoviolaceus OPC-520.
Tsujibo H, Endo H, Miyamoto K, Inamori Y. Tsujibo H, et al. Biosci Biotechnol Biochem. 1995 Jan;59(1):145-6. doi: 10.1271/bbb.59.145. Biosci Biotechnol Biochem. 1995. PMID: 7765967
Cited by
- Wolbachia depletion blocks transmission of lymphatic filariasis by preventing chitinase-dependent parasite exsheathment.
Quek S, Cook DAN, Wu Y, Marriott AE, Steven A, Johnston KL, Ford L, Archer J, Hemingway J, Ward SA, Wagstaff SC, Turner JD, Taylor MJ. Quek S, et al. Proc Natl Acad Sci U S A. 2022 Apr 12;119(15):e2120003119. doi: 10.1073/pnas.2120003119. Epub 2022 Apr 4. Proc Natl Acad Sci U S A. 2022. PMID: 35377795 Free PMC article. - Microbial chitinases: properties, enhancement and potential applications.
Gomaa EZ. Gomaa EZ. Protoplasma. 2021 Jul;258(4):695-710. doi: 10.1007/s00709-021-01612-6. Epub 2021 Jan 22. Protoplasma. 2021. PMID: 33483852 Review. - Microbial chitinases: properties, current state and biotechnological applications.
Le B, Yang SH. Le B, et al. World J Microbiol Biotechnol. 2019 Sep 6;35(9):144. doi: 10.1007/s11274-019-2721-y. World J Microbiol Biotechnol. 2019. PMID: 31493195 Review. - Development and optimization of a high-throughput screening method utilizing Ancylostoma ceylanicum egg hatching to identify novel anthelmintics.
Abriola L, Hoyer D, Caffrey CR, Williams DL, Yoshino TP, Vermeire JJ. Abriola L, et al. PLoS One. 2019 Jun 3;14(6):e0217019. doi: 10.1371/journal.pone.0217019. eCollection 2019. PLoS One. 2019. PMID: 31158236 Free PMC article. - A novel small molecule that disrupts a key event during the oocyte-to-embryo transition in C. elegans.
Weicksel SE, Mahadav A, Moyle M, Cipriani PG, Kudron M, Pincus Z, Bahmanyar S, Abriola L, Merkel J, Gutwein M, Fernandez AG, Piano F, Gunsalus KC, Reinke V. Weicksel SE, et al. Development. 2016 Oct 1;143(19):3540-3548. doi: 10.1242/dev.140046. Epub 2016 Aug 10. Development. 2016. PMID: 27510972 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
Research Materials