Research Article | OPEN ACCESS
Purification Crude Extracellular Protease of Saccharomycopsis fibuligera strain R64 Isolated from Tape' Indonesian Fermented Food
1L.B. Roostita, 1W.S. Putranto, 2C. Charoenchai, 1E. Wulandari and 3L.U. Gemilang
1Faculty of Animal Husbandry, University of Padjadjaran, West Java, Indonesia
2Faculty of Home Economics Technology, Rajamangala Univ. of Technology Thanyaburi, Thailand
3Graduate Program of Environmental Science, University of Padjadjaran, West Java, Indonesia
Advance Journal of Food Science and Technology 2013 8:1002-1004
Received: March 04, 2013 | Accepted: April 02, 2013 | Published: August 05, 2013
Abstract
The research aims to purify crude extracellular protease produced by Saccharomycopsis fibuligera strain R64 that isolated from tape’ Indonesian fermented food. Saccharomycopsis fibuligera strain R64 was tested on Malt Extract Agar (MEA) that added 3% skim milk powder to discover the proteolytic activity. Extracellular protease purified with ammonium sulfate precipitation, dialysis and gel filtration chromatography with Sephadex G-100. Bradford method used to measure protein content, protease activity measured with Walter method and SDS-PAGE used to measure the molecule weight of protease. Results shown that isolated yeasts give 2-5 mm of clear zones on MEA that added with skim milk powder. Precipitations with 45% ammonium sulfate give the best protein content of 5.05 mg/mL and protease activity of 0.051 U/mL. After dialysis the protein content the protease activity were increased to 5.57 mg/mL and 0.0631 U/mL. Moreover, the gel filtration chromatography with sephadex G-100 yields 0.023 U/mL proteolytic activities, 2.1 mg/mL protein contents with 97.4 kD protease molecular weight.
Keywords:
Crude extracellular protease, purification, Saccharomycopsis fibuligera strain R64, tape,
Competing interests
The authors have no competing interests.
Open Access Policy
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Copyright
The authors have no competing interests.
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ISSN (Online): 2042-4876
ISSN (Print): 2042-4868 |
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