Cloning and expression analysis of the chitinase gene Ifu-chit2 from Isaria fumosorosea

Abstract

Entomopathogenic fungi can produce a series of chitinases, some of which function synergistically with proteases and other hydrolytic enzymes to degrade the insect cuticle. In the present study, the chitinase gene Ifu-chit2 from Isaria fumosorosea was investigated. The Ifu-chit2 gene is 1,435-bp long, interrupted by three short introns, and encodes a predicted protein of 423 amino acids with a 22 residue signal peptide. The predicted Ifu-Chit2 protein is highly homologous to Beauveria bassiana chitinase Bbchit2 and belongs to the glycohydrolase family 18. Ifu-Chit2 was expressed in Escherichia coli to verify chitinase activity, and the recombinant enzyme exhibited activity with a colloidal chitin substrate. Furthermore, the expression profiles of Ifu-chit2 were analyzed at different induction times under in vivo conditions. Quantitative real-time PCR analysis revealed that Ifu-chit2 expression peaked at two days post-induction. The expression of chitinase Ifu-chit2 in vivo suggests that the chitinase may play a role in the early stage of pathogenesis.

Isolation, Characterization, and Insecticidal Activity of Chitinase Gene (chiA) From Aspergillus Niger (EM77)

“…Based on the similarity percentages of amino acid sequences, most chitinase genes that have been characterized to date are classified into two families of glycosyl hydrolases (family18 and 19) (Meng et al, 2015). Thus, the deduced amino acid Isolation, Characterization, and Insecticidal Activity of Chitinase Gene (chiA) 119 sequence of chitinase (chiA) of the isolate EM77 has included the characteristic substrate-binding site and the catalytic motifs cited by Vocadlo and Davies (2008).…”

Chitinolytic fungi from the Birjand plain of Southern Khorasan Province in Eastern Iran

“…The presence of chitinase enzyme genes was investigated by amplification of extracted DNA from fungal isolates by polymerase chain reaction (PCR) using the Chit2 degenerative primer set (forward primer: 5'-TCCATYGGNGGNTGGACNTG-3' and reverse primer: 5'-GCRSWNGCYTCCCARAACAT-3') and DECH degenerative primer set (forward primer: 5'-TCCCARAYHCCRTTCTCCCA-3', and reverse primer: 5'-AAYYTBATGGCYTAYGACT-3') [36]. DNA was extracted from the fungal isolates using 600-800 μL of cetyltrimethyl ammonium bromide (CTAB) 2× extraction buffer and 1 μL of proteinase K (Cinnagen Co., Iran) added to the fungal cells and incubated 65-70°C for 60 min.…”

Extraction and Toxicity Assay of Mycotoxin from Entomopathogenic Fungi Isolate of Kusuma Agrowisata Orchard Batu, Jawa Timur, Indonesia

“…There are still other mycotoxin from entomopathogenic fungi that has not been identified by name as researched by Meng et al [28], with a size of about 50 kDa derived from Beauveria sp.…”