DOI QR코드

DOI QR Code

Optimal Culture Conditions for Production of Subtilisin-like Protease Inhibitor from Streptomyces thermocarboxydus C12

Streptomyces thermocarboxydus C12에서 Subtilisin-like Protease Inhibitor 생산을 위한 최적배양조건

  • Kang, Sung-Il (Dept. of Biotechnology & Bioengineering, Pukyong National University) ;
  • Jang, Young-Boo (Dept. of Biotechnology & Bioengineering, Pukyong National University) ;
  • Choi, Gyeong-Lim (Division of Marine Bioscience/Institue of Marine Industry, Gyeongsang National University) ;
  • Choi, Byeong-Dae (Division of Marine Bioscience/Institue of Marine Industry, Gyeongsang National University) ;
  • Kong, Jai-Yul (Dept. of Biotechnology & Bioengineering, Pukyong National University) ;
  • Choi, Yeung-Joon (Division of Marine Bioscience/Institue of Marine Industry, Gyeongsang National University)
  • 강성일 (부경대학교 생물공학과) ;
  • 장영부 (부경대학교 생물공학과) ;
  • 최경임 (경상대학교 해양식품생명공학과) ;
  • 최병대 (경상대학교 해양식품생명공학과) ;
  • 공재열 (부경대학교 생물공학과) ;
  • 최영준 (경상대학교 해양식품생명공학과)
  • Published : 2008.03.31

Abstract

The objective of this paper was to investigate optimal culture conditions for the production of an inhibitor against subtilisin-like protease from Streptomyces thermocarboxydus (S. thermocarboxydus) C12 isolated from sediments of Gwangyang coast. The optimal temperature and initial pH for the production of subtilisin-like protease inhibitor were $40^{\circ}C$ and pH 8.0, respectively. Inhibition activities were high for galactose, glucose and fructose. The best carbon source and its concentration were galactose and 1.6% (w/v), respectively. Inhibition activities were also high in medium containing polypeptone, proteose and peptone. Optimal nitrogen source and concentration were protease peptone and 0.5% (w/v), respectively. Optimal concentrations for inhibitor production were 1% (w/v) for NaCl and 1 mM LiCl for metal salts. The subtilisin-like protease inhibitor from S. thermocarboxydus C12 showed a maximum inhibitor activity after cultivation for 84 h under the optimized medium.

광양만의 해안 뻘에서 protease 저해력이 우수한 균주 S. thermocarboxydus C12를 분리하여 최적의 저해제 생산 조건을 조사하였다. 최대 저해활성을 보이는 온도와 초기 pH는 각각 $40^{\circ}C$와 8.0이었다. 저해제 생산에 미치는 탄소원의 영향을 검토한 결과, 단당류인 galactose, glucose, fructose와 다당류인 starch에서 높은 저해활성을 보였으며, 최적의 탄소원과 농도는 각각 galactose와 1.6%(w/v)였다. 질소원의 경우 복합배지인 polypeptone과 proteose peptone에서 가장 높은 저해활성이 나타났으며, 최적의 농도와 질소원은 0.5%(w/v)와 proteose peptone이었다. 최적의 NaCl 농도와 금속이온은 각각 1%(w/v)와 1 mM LiCl이었다. 선정된 최적배양조건에서 균주를 배양한 결과, 84시간 동안 배양 시 저해활성이 최대인 것으로 나타났다.

Keywords

References

  1. Perlmann GE, Lorand L. 1970. Proteolytic enzymes. In Methods in Enzymology. Academic press, New York, USA. Vol XIX, p 807-889
  2. Umezawa H. 1972. Enzyme inhibitors of microbial origin. University of Tokyo Press, Tokyo, Japan. p 1-114
  3. Bode W, Huber R. 1992. Natural protein proteinase inhibitors and their interaction with proteinases. Eur J Biochem 204: 433-451 https://doi.org/10.1111/j.1432-1033.1992.tb16654.x
  4. Terashita T, Kono M, Murao S. 1980. Promoting effect of S-PI on fruiting of Lentinus edodes. Trans Mycol Soc Jpn 21: 137-140
  5. Garcia-Carreno FL. 1996. Proteinase inhibitors. Trends Food Sci Technol 7: 197-204 https://doi.org/10.1016/0924-2244(96)10023-6
  6. Demuth HU. 1990. Recent developments in inhibiting cysteine and serine proteases. J Enzym Inhibition 3: 249-278 https://doi.org/10.3109/14756369009030375
  7. Aoyagi T, Takeuchi T, Matsuzaki A, Kawamura K, Kondo S, Hamada M, Maeda K, Umezaua H. 1969. Leupeptins, new protease inhibitors from Actinomyces. J Antibiot 22: 283-286 https://doi.org/10.7164/antibiotics.22.283
  8. Umezawa S, Tatsuta K, Fujimoto K, Tsuchiya T, Umezawa H. 1972. Structure of antipain, a new Sakaguchi-positive product of Streptomcyes. J Antibiot 25: 267-270 https://doi.org/10.7164/antibiotics.25.267
  9. Tatsuta K, Mikami N, Fujimoto K, Umezawa S, Umezawa H. 1973. The structure of chymostatin, a chymotrypsin inhibitor. J Antibiot 26: 625-646 https://doi.org/10.7164/antibiotics.26.625
  10. Kunimoto S, Aoyagi T, Morishima H, Takeuchi T, Umezawa H. 1972. Mechanism of inhibition of pepsin by pepstatin. J Antibiot 25: 251-255 https://doi.org/10.7164/antibiotics.25.251
  11. Murao S, Sato S. 1972. S-SI, a new alkaline protease inhibitor from Streptomyces albogriseolus S-3253. Agric Biol Chem 36: 160-163 https://doi.org/10.1271/bbb1961.36.160
  12. Kourteva Y, Boteva R. 1989. A novel extracellular subtilisin inhibitor produced by a Streptomyces sp. FEB 247: 468-470 https://doi.org/10.1016/0014-5793(89)81392-4
  13. Terabe M, Kojima S, Taguchi S, Momose H, Miura K-I. 1996. New subtilisin-trypsin inhibitors produced by Streptomyces: primary structure and their relationship to other proteinase inhibitors from Streptomyces. Biochim Biophys Acta 1292: 233-240 https://doi.org/10.1016/0167-4838(95)00207-3
  14. Taguchi S, Kojima S, Terabe M, Kumazawa Y, Kohriyama H, Suzuki M, Miura K-I, Momose H. 1997. Molecular phylogenetic characterization of Streptomyces protease inhibitor family. J Mol Evol 44: 542-551 https://doi.org/10.1007/PL00006178
  15. Kang S-I. 2007. Identification and purification properties of subtilisin-like proteinase inhibitor isolated from the marine bacterium. PhD Dissertation. Pukyong National University, Busan
  16. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275
  17. Moore S, Stein WH. 1948. Photometric ninhydrin method for use in the chromatography of amino acids. J Biol Chem 176: 367-388
  18. Kang SI, Kim YM, Jang YB, Lim DJ, Kong JY. 2004. The optimal culture condition for the collagenolytic protease production from Vibrio vulnificus CYK279H. Korean J Biotechnol Bioeng 19: 295-300
  19. Choi HJ, Jung MJ, Jeong YK. 2002. Optimization of the production of an immunostimulant from a marine bacterium. Korean J Life Sci 12: 759-764 https://doi.org/10.5352/JLS.2002.12.6.759
  20. Murao S, Sato S, Muto N. 1972. Isolation of alkaline protease inhibitor producing microorganisms. Agric Biol Chem 36: 1737-1744 https://doi.org/10.1271/bbb1961.36.1737
  21. Tsuchiya K, Kimura T. 1978. Production of trypsin inhibitor by a Cephalosporium sp. Appl Environ Microbiol 35: 631-635
  22. Fukuhara KI, Katsura M, Murao S. 1982. Purification and some properties of Talopeptin (MK-1), a novel proteinase inhibitor produced by Streptomyces mozunensis MK-23. Agric Biol Chem 46: 1707-1710 https://doi.org/10.1271/bbb1961.46.1707
  23. Park SK, Sung NK, Lee SW. 1989. Production and purification of pepsin inhibitor from Actinomyces GF 155-2. Korean J Appl Microbiol Bioeng 17: 121-125
  24. Murao S, Watanabe T. 1978. Isolation and identification of microorganism, producing microbial alkaline proteinase inhibitor (MAPI). Agric Biol Chem 42: 2209-2215 https://doi.org/10.1271/bbb1961.42.2209
  25. Kim IS, Kim HT, Lee HS, Lee KJ. 1991. Protease inhibitor production using Streptomyces sp. SMF13. J Microbiol Biotechnol 1: 288-292
  26. Pandhare J, Zog K, Deshpande VV. 2002. Differential stability of alkaline protease inhibitors from actinomyces: effect of various additives on thermostability. Bioresour Technol 84: 165-169 https://doi.org/10.1016/S0960-8524(02)00025-1
  27. Aoyagi T. 1990. Small molecular protease inhibitors and their biological effects. In Biochemistry of Peptide Antibiotics. Kleinkauf H, Dohren H, eds. Walter de Gruyter, Berlin. p 312-363
  28. Uo ME, Joo DS, Cho SY. 2006. Screening and cultivation characteristics of alginate degrading bacteria. J Korean Soc Food Sci Nutr 35: 109-114 https://doi.org/10.3746/jkfn.2006.35.1.109
  29. 안영길. 1993. 단백질의 변성과 재생, 효소화학. 청문각, 서울. p 118-125