Cloning, isolation, and characterization of novel chitinase-producing bacterial strain UM01 (Myxococcus fulvus)

Abstract

Background: Chitin is an important biopolymer next to cellulose, extracted in the present study. The exoskeleton of marine bycatch brachyuran crabs, namely Calappa lophos, Dromia dehaani, Dorippe facchino and also from stomatopod Squilla spp. were used to extract chitin through fermentation methods by employing two bacterial strains such as Pseudomonas aeruginosa, Serratia marcescens. The yield of chitin was 44.24%, 37.45%, 11.56% and 27.24% in C. lophos, D. dehaani, D. facchino and Squilla spp. respectively. FT-IR spectra of the produced chitin exhibit peaks which is more or less coherent to that of standard chitin which is further analysed by Scanning Electron Microscope. The quality of produced chitin was assessed through moisture, protein, ash and lipid content analysis ensured that chitin obtained from trash crustaceans are on par with that of standard chitin. Results: A total of 10 samples were collected from different areas of Jiangsu China for screening of chitinaseproducing bacteria. Based on the clearance zone, two of the best samples were chosen for further study. 16S rRNA sequence analysis showed that this strain belongs to genus Myxococcus and species Myxococcus fulvus. Phylogenetic analysis was performed and it shows strain UM01 is a novel bacterial strain. UM01 isolate shows maximum chitinase production at 35°C and 8 pH. Among all, these colloidal chitins were found to be the best for chitinase production. Three chitinase-producing genes were identified and sequenced by using degenerative plasmid. UMCda gene (chitin disaccharide deacetylase) was cloned into E. coli DH5a by using PET-28a vector, and antagonistic activity was examined against T. reesei. Conclusion: To our knowledge, this is the earliest study report to gene cloning and identification of the chitinase gene in Myxococcus fulvus. Chitinase plays a key role in decomposition and utilization of chitin as a raw material. This research indicates that Myxococcus fulvus UM01 strain is a novel myxobacteria strain and can produce large amounts of chitinase within a short time. The UMCda gene cloned into E. coli DH5a showed a promising effect as antifungal activity. In overall findings, the specific strain UM01 has endowed properties of bioconversation of waste chitin and other biological applications.

A Broad-Specificity Chitinase from Penicillium oxalicum k10 Exhibits Antifungal Activity and Biodegradation Properties of Chitin

“…Chitinases from Acremonium sp. YS2-2 [ 8 ], Trichoderma asperellum PQ34 [ 21 ], Myxococcus fulvus UM01 [ 22 ], and Achromobacter xylosoxidans [ 23 ] were induced in response to 1% colloidal chitin. The maximum chitinase yield of Trichoderma viride [ 24 ], Streptomyces pratensis KLSL55 [ 25 ], and Hydrogenophilus hirschii B-DZ44 [ 26 ] were obtained at 1.4%, 1.5%, and 2% of colloidal chitin, respectively.…”

Western Kenyan Anopheles gambiae showing intense permethrin resistance harbour distinct microbiota

“…On the other hand, the genus Myxococcus was only detected in susceptible mosquitoes. This bacterial genus is known to be predatory on other bacteria [63], chitinase-producing [64], capable of producing various bioactive antifungal agents [65], and inhibitors of cellular respiration [66]. However, their association with mosquito physiology or insecticide susceptibility has not yet been described.…”

In silico analysis of promoter region and regulatory elements of mitogenome co-expressed trn gene clusters encoding for bio-pesticide in entomopathogenic fungus, Metarhizium anisopliae: strain ME1

“…Besides to this, studies have been conducted to understand the molecular level infection pathways of insects by examining M. anisopliae species, for example: laboratory tests using M. anisopliae against ticks [7], controlling rice leaf roller [8], and mortality of migratory locust [9]. There is evolving evidence that insect pathogenic fungi are unique in being able to infect across the insect cuticle [10] and attack the insect by invading cuticle layer [11,12]. In contrast, other organisms like viruses and bacteria attack their host after ingestion [5].…”