Metabolic pathway for poly(3-hydroxybutyrate-co-3-hydroxyvalerate) formation in Nocardia corallina: inactivation of mutB by chromosomal integration of a kanamycin resistance gene.

Applied and Environmental Microbiology
H E Valentin, D Dennis

Abstract

The gene encoding the large subunit of the methylmalonyl-coenzyme A (CoA) mutase in Nocardia corallina (mutBNc) was cloned. A 4.3-kbp BamHI fragment containing almost the entire mutBNc was identified by Southern hybridization experiments employing a digoxigenin-labeled probe deduced from mutB of Streptomyces cinnamonensis, mutBNc was interrupted by insertion of a kanamycin resistance gene block (mutB::kan or mutB::neo) and introduced into N. corallina to obtain mutB-negative strains by homologous recombination. Four of sixteen kanamycin-resistant clones occurred via double-crossover events and harbored only the interrupted mutBNc. These exhibited no growth on odd-chain fatty acids in the presence of kanamycin but exhibited wild-type growth on even-chain fatty acids, glucose, and succinate. Whereas the wild type of N. corallina accumulates a copolyester of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) containing more than 60 mol% 3HV from most carbon sources, mutB-negative strains accumulated poly(3HB-co-3HV) containing only 2 to 6 mol% 3HV. Methylmalonyl-CoA mutase activity was not found in these clones. Therefore, this study provides strong evidence that the majority of 3HV units in poly(3HB-co-3HV) accumulated by N. cor...Continue Reading

References

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Apr 1, 1992·Applied and Environmental Microbiology·S SlaterD Dennis
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Citations

Dec 22, 2009·Applied Microbiology and Biotechnology·Jorge QuillaguamánRajni Hatti-Kaul
Sep 29, 2004·FEMS Microbiology Reviews·Volker GürtlerRobert Seviour
Feb 1, 2017·Standards in Genomic Sciences·Nyok-Sean LauAbdullah Al-Ashraf Amirul
May 7, 2019·Expert Review of Medical Devices·Enas ElmowafyVincenzo Guarino
Nov 10, 1998·Trends in Biotechnology·A Steinbüchel, B Füchtenbusch
Mar 6, 1999·Microbiology and Molecular Biology Reviews : MMBR·L L Madison, G W Huisman
Jun 17, 2021·Essays in Biochemistry·Dillirani NagarajanJo-Shu Chang
Oct 20, 2004·Metabolic Engineering·Andrew R ReevesJ Mark Weber

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