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| Title: | A solvent stable metalloprotease produced by Bacillus sp. TKU004 and its application in the deproteinization of squid pen for β-chitin preparation |
| Authors: | Wang, San-lang;Chern, Ming-kai;Chen, Yau-hung |
| Contributors: | 淡江大學生命科學研究所 |
| Keywords: | Bacillus;Protease;Squid pen;Chitin;Deproteinization |
| Date: | 2006-08 |
| Issue Date: | 2013-02-27 09:39:12 (UTC+8) |
| Publisher: | Philadelphia: Elsevier Inc. |
| Abstract: | A protease-producing bacterium was isolated and identified as Bacillus sp. TKU004. It thus can be used for deproteinization of squid pen in the preparation of β-chitin. The optimized condition for protease production was found when the culture was shaken at 30 °C for 4 days in 100 mL of medium containing 2% squid pen powder (SPP) (w/v), 0.1% K2HPO4, and 0.05% MgSO4. Under such condition, both the production of protease by Bacillus sp. TKU004 and the resulted protein removal of squid pen attained the optimum. They were 0.065 U/mL and 73%, respectively. An extracellular protease was purified from the culture supernatant of Bacillus sp. TKU004. The molecular weight of TKU004 protease was 27 and 57 kDa assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively. The optimum pH, optimum temperature, pH stability, thermal stability, Km, Vmax, and activation energy of TKU004 protease were 6–8, 60 °C, pH 5–8, 50 °C, 2.98 mg/mL, 0.14 U/mL, 8.31 J/(mol K), respectively. It was concluded that TKU004 protease is a Zn-containing metalloenzyme, and its dimeric structure contains at least one disulfide bond. The unique characteristic of TKU004 protease is that it retained more than 60% of its original activity after preincubation for 10 days at 25 °C in the presence of tested organic solvents (25%, v/v). This is also the first report of a strain being able to use squid pen as the sole carbon/nitrogen source for proteases production and being used for the reclamation of β-chitin from squid pen. |
| Relation: | Enzyme and Microbial Technology 39(4), p.724-731 |
| DOI: | 10.1016/j.enzmictec.2005.12.007 |
| Appears in Collections: | [生命科學研究所] 期刊論文
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