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"0022-5142", "eIssn" : "1097-0010", "reviewType" : "REVIEWED", "noIF" : false, "sciIndexed" : true, "scopusIndexed" : true, "lang" : "FOREIGN", "hungarian" : false, "published" : true, "oldId" : 2853, "snippet" : true }, "volume" : "100", "issue" : "7", "firstPage" : "3013", "lastPage" : "3023", "firstPageOrInternalIdForSort" : "3013", "pageLength" : 11, "publishedYear" : 2020, "abstractText" : "BACKGROUND Mesophilic alpha-amylases function effectively at low temperatures with high rates of catalysis and require less energy for starch hydrolysis. Bacillus amyloliquefaciens is an essential producer of mesophilic alpha-amylases. However, because of the existence of the restriction-modification system, introducing exogenous DNAs into wild-type B. amyloliquefaciens is especially tricky.RESULTS alpha-Amylase producer B. amyloliquefaciens strain Z3 was screened and used as host for endogenous alpha-amylase gene expression. In vitro methylation was performed in recombinant plasmid pWB980-amyZ3. With the in vitro methylation, the transformation efficiency was increased to 0.96 x 10(2) colony-forming units mu g(-1) plasmid DNA. A positive transformant BAZ3-16 with the highest alpha-amylase secreting capacity was chosen for further experiments. The alpha-amylase activity of strain BAZ3-16 reached 288.70 +/- 16.15 U mL(-1) in the flask and 386.03 +/- 16.25 U mL(-1) in the 5-L stirred-tank fermenter, respectively. The Bacillus amyloliquefaciens Z3 expression system shows excellent genetic stability and high-level extracellular production of the target protein. Moreover, the synergistic interaction of AmyZ3 with amyloglucosidase was determined during the hydrolysis of raw starch. The hydrolysis degree reached 92.34 +/- 3.41% for 100 g L-1 raw corn starch and 81.30 +/- 2.92% for 100 g L-1 raw cassava starch after 24 h, respectively.CONCLUSION Methylation of the plasmid DNA removes a substantial barrier for transformation of B. amyloliquefaciens strain Z3. Furthermore, the exceptional ability to hydrolyze starch makes alpha-amylase AmyZ3 and strain BAZ3-16 valuable in the starch industry. 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