Abstract
Hybrid β-glucanase genes were constructed by the reciprocal exchange of the two halves of the isolated β-glucanase genes from Bacillus amyloliquefaciens and B. macerans. The β-glucanase hybrid enzyme 1 (H1) contains the 107 amino-terminal residues of mature B. amyloliquefaciens β-glucanase and the 107 carboxyl-terminal amino acid residues of B. macerans β-glucanase. The reciprocal β-glucanase hybrid enzyme 2 (H2) consists of the 105 amino-terminal residues from the B. macerans enzyme and the carboxyl-terminal 107 amino acids from B. amyloliquefaciens. The biochemical properties of the two hybrid enzymes differ significantly from each other as well as from both parental β-glucanases.
Hybrid β-glucanase H1 exhibits increased thermostability in comparison to other β-glucanases, especially in an acidic environment. This hybrid enzyme has maximum activity between pH 5.6 and 6.6, whereas the pH-optimum for enzymatic activity of B. amyloliquefaciens β-glucanase was found to be at pH 6 to 7 and for B. macerans at pH 6.0 to 7.5 Hybrid enzyme 1 being more heat stable than both parental enzymes represents a case of intragenic heterosis.
Hybrid β-glucanase 2 (H2) was found to be more thermolabile than the naturally occurring β-glucanases it was derived from and the pH-optimum for enzymatic activity was determined to be between pH 7 and pH 8.
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Abbreviations
- aa:
-
amino acids
- amp:
-
ampicillin
- bgl:
-
β-glucanase gene
- β-glucan:
-
(1–3, 1–4)-β-d-glucan
- PAGE:
-
polyacrylamide gel electrophoresis
- SDS:
-
sodium dodecyl sulfate
- tet:
-
tetracycline
- SP:
-
signal peptide
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Borriss, R., Olsen, O., Thomsen, K.K. et al. Hybrid bacillus endo-(1–3, 1–4)-β-glucanases: Construction of recombinant genes and molecular properties of the gene products. Carlsberg Res. Commun. 54, 41 (1989). https://doi.org/10.1007/BF02907584
DOI: https://doi.org/10.1007/BF02907584