Abstract
Alpha amylase gene from Bacillus licheniformis was mutated by site-directed mutagenesis to improve its acid stability. The mutant gene was expression in Bacillus subtilis under the control of the promoter of sacB gene which was followed by either the α-amylase leader peptide of Bacillus licheniformis or the signal peptide sequence of sacB gene of Bacillus subtilis. Both peptides efficiently directed the secretion of α-amylase from the recombinant B. subtilis cells. The extracellular α-amylase activities in two recombinants were 1001 and 2012 U ml−1, respectively. The purity of the recombinant product was confirmed by SDS-PAGE.
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References
W Aehle O Misset (1999) Enzymes for industrial applications HJ Rehm G Reed (Eds) Biotechnology, Vol. 5a EditionNumber2 Wiley-VCH Germany 191–198
WD Crabb JK Shetty (1999) ArticleTitleCommodity scale production of sugars from starches Curr. Opin. Microbiol. 2 252–256 Occurrence Handle10.1016/S1369-5274(99)80044-7 Occurrence Handle10383865
RH Doi SL Wong F Kawamura (1998) ArticleTitlePotential use of Bacillus subtilis for secretion and production of foreign proteins Trends Biotechnol. 4 232–235 Occurrence Handle10.1016/0167-7799(86)90116-2
D Dubnau TJ Gryczan S Contente (1978) ArticleTitleCharacterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis J. Bacteriol. 134 318–329 Occurrence Handle418061
SD Ehrlich (1977) ArticleTitleReplication and expression of plasmids from Staphylococcus aureus in Bacillus subtilis Proc. Natl. Acad. Sci. USA 70 1680–1682
SD Ehrlich B Niaudet B Michel (1982) ArticleTitleUse of plasmids from Staphylococcus aureus for cloning of DNA in Bacillus subtilis Curr. Top. Microbiol. Immunol. 96 19–29 Occurrence Handle6276091
DV Goeddle (1990) ArticleTitleExpression in B. subtilis Meth. Enzymol. 85 199–228
SN Ho HD Hunt RM Horton JK Pullen LR Pease (1989) ArticleTitleSite-directed mutagenesis by overlap extension using the polymerase chain reaction Gene 77 51–59 Occurrence Handle10.1016/0378-1119(89)90358-2 Occurrence Handle2744487
K Ikawa H Araki Y Tsujino Y Hayashi K Igarashi Y Hatada H Hagihara T Ozawa K Ozaki T Kobayashi S Ito (1998) ArticleTitleHyperexpression of the gene for a Bacillus alpha-amylase in Bacillus subtilis cells: enzymatic properties and crystallization of the recombinant enzyme Biosci. Biotechnol. Biochem. 62 1720–1725 Occurrence Handle10.1271/bbb.62.1720 Occurrence Handle9805372
JJ Lee KS Hahm KY Lee ST Lee (1997) ArticleTitleCharacterization of an endoxylanase produced by an isolated strain of Bacillus sp J. Microbiol. Biotechnol. 7 114–120
LL Lin WH Hsu (1997) ArticleTitleLactose-induced expression of Bacillus sp. TS-23 amylase gene in E. coli regulated by a T7 promoter Lett. Appl. Microbiol. 24 365–368 Occurrence Handle10.1046/j.1472-765X.1997.00146.x Occurrence Handle9172444
A Nakayama K Ando K Kawamura I Mita K Fukazawa M Hori M Honjo Y Furutani (1988) ArticleTitleEfficient secretion of the authentic mature human growth hormone by Bacillus subtilis J. Biotechnol. 8 123–134 Occurrence Handle10.1016/0168-1656(88)90074-0
CS Park CC Chang JY Kim (1997) ArticleTitleExpression, secretion, and processing of rice a-amylase in the Yarrowia lipolytica J. Biol. Chem. 272 6876–6881 Occurrence Handle10.1074/jbc.272.11.6876 Occurrence Handle9054373
J Sambrook EF Fritsch T Maniatis (1989) Molecular Cloning, A Laboratory Manual New York Cold Spring Harbor
F Sanger S Nicklen AR Coulson (1977) ArticleTitleDNA sequencing with chain-terminating inhibitors Proc. Natl. Acad. Sci. USA 74 5463–5467 Occurrence Handle271968
A Shaw R Bott (1996) ArticleTitleEngineering enzymes for stability Curr. Opin. Biotechnol. 6 546–550
Takazaki Y, Hirose S, (1996) Acid-stable and thermostable α-amylase gene. Japanese Patent 8-289788 (in Japanese)
JG Thomas F Baneyx (1996) ArticleTitleProtein misfolding and inclusion body formation in recombinant Escherichia coli cells overexpressing heat-shock proteins J. Biol. Chem. 271 11141–11147 Occurrence Handle10.1074/jbc.271.19.11141 Occurrence Handle8626659
SL Wong F Kawamura RH Doi (1986) ArticleTitleUse of the Bacillus subtilis subtilisin signal peptide for efficient secretion of TEM β-lactamase during growth J. Bacteriol. 168 1005–1009 Occurrence Handle3096954
Q Wu JX Luo BN Xu WQ Li (2001) ArticleTitleConstruction of high level inducible expression-secretion system of Bacillus subtilis Evol. Nat. Sci. 11 40–46
W Xu-chu L Wilson Louise Tran W Sui-lam (1991) ArticleTitleEngineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases J Bacteriol. 173 4952–4958 Occurrence Handle1907264
R Ye JH Kim BG Kim S Szarka E Sihota SL Wong (1999) ArticleTitleHigh level production of intact, biologically active staphylokinase from Bacillus subtilis Biotechnol. Bioeng. 62 87–96 Occurrence Handle10.1002/(SICI)1097-0290(19990105)62:1<87::AID-BIT10>3.0.CO;2-I Occurrence Handle10099516
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Heng, C., Chen, Z., Du, L. et al. Expression and Secretion of an Acid-Stable α-Amylase Gene in Bacillus Subtilis by SacB Promoter and Signal Peptide. Biotechnol Lett 27, 1731–1737 (2005). https://doi.org/10.1007/s10529-005-2743-4
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DOI: https://doi.org/10.1007/s10529-005-2743-4