title: Molecular characterisation of Bacillus chitinase for bioconversion of chitin waste.
abstract: In this work chitin was extracted chemically from shrimp shells. Seventeen Bacillus isolates were screened for chitinolytic activity. The chitinolytic strains of Bt. were screened at different temperatures and pHs for their hydrolytic potentials. By using a pair of specific primers, endochitinase gene was amplified from SBS Bt-5 strain through PCR, and then cloned into pTZ57 TA cloning vector and transferred in Escherichia coli DH5α strain. The sequenced gene (GenBank Accession No: HE995800) consists of 2031 nucleotides capable of encoding 676 residues. The protein consisted of three functional domains with a calculated molecular mass of 74.53 kDa and a pI value of 5.83. The amino acid sequence of chi gene showed 99% similarity to the genes of Bt MR11 endochitinase, Bt serovar kurstaki chitinase (kchi), Bt strain MR21 endochitinase and Bacillus cereus B4264.
authors.affiliation: a Department of Biotechnology , Lahore College for Women University , Lahore , Pakistan.
authors.forename: Faiza
authors.initials: F
authors.lastname: Saleem
authors.affiliation: a Department of Biotechnology , Lahore College for Women University , Lahore , Pakistan.
authors.forename: Uzma
authors.initials: U
authors.lastname: Nisar
authors.affiliation: a Department of Biotechnology , Lahore College for Women University , Lahore , Pakistan.
authors.forename: Aisha
authors.initials: A
authors.lastname: Younas
authors.affiliation: b Department of Zoology , University of the Punjab , Lahore , Pakistan.
authors.forename: Faiza
authors.initials: F
authors.lastname: Jabeen
authors.affiliation: b Department of Zoology , University of the Punjab , Lahore , Pakistan.
authors.forename: Javed Iqbal
authors.initials: JI
authors.lastname: Qazi
authors.affiliation: b Department of Zoology , University of the Punjab , Lahore , Pakistan.
authors.forename: Nazia
authors.initials: N
authors.lastname: Khursheed
authors.affiliation: a Department of Biotechnology , Lahore College for Women University , Lahore , Pakistan.
authors.forename: Neelma
authors.initials: N
authors.lastname: Munir
authors.affiliation: a Department of Biotechnology , Lahore College for Women University , Lahore , Pakistan.
authors.forename: Shagufta
authors.initials: S
authors.lastname: Naz
authors.affiliation: c School of Biological Sciences, University of the Punjab , Lahore , Pakistan.
authors.forename: Abdul Rauf
authors.initials: AR
authors.lastname: Shakoori
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D002686:Chitin / Q000378:metabolism*; D002688:Chitinases / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D002686:Chitin; D002688:Chitinases
keywords: N-acetyl-d-glucosamine; chitin; chitinolytic bacteria; endochitinase; shrimp shells