α-Glucosidase inhibitory activity and protease characteristics produced by Bacillus amyloliquefaciens

Bacillus amyloliquefaciens로부터 생산된 protease 특성 및 α-glucosidase 저해활성

  • Lee, Rea-Hyun (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Yang, Su-Jin (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Hwang, Tae-Young (Department of Food Science and Technology, Jungwon University) ;
  • Chung, Shin-Kyo (School of Food Science and Biotechnology, Kyungpook National University) ;
  • Hong, Joo-Heon (Department of Food Science and Technology, Catholic University of Daegu)
  • 이래현 (대구가톨릭대학교 식품공학전공) ;
  • 양수진 (대구가톨릭대학교 식품공학전공) ;
  • 황태영 (중원대학교 식품공학과) ;
  • 정신교 (경북대학교 식품공학부) ;
  • 홍주헌 (대구가톨릭대학교 식품공학전공)
  • Received : 2015.07.27
  • Accepted : 2015.10.07
  • Published : 2015.10.30


In this study, three GRAS (generally recognized as safety) strain was isolated from Doenjang and Cheonggukjang and identified as a protease-producing microorganism, following the appearance of a clear zone around its colony when cultured on a medium containing skim milk. Based on an analysis of the nucleotide sequence of 16S ribosomal RNA, the strains wereas identified as Bacillus amyloliquefaciens and wereas therefore named Bacillus amyloliquefaciens CDD5, Bacillus amyloliquefaciens CPD4, and Bacillus amyloliquefaciens CGD3. Here, we analyzed the protease and ${\alpha}$-glucosidase inhibitory activities of the three B. amyloliquefaciens strains. Among the isolated strains, B. amyloliquefaciens CGD3 exhibited the highest protease activity (9.21 U/mL, 24 hr). The protease activities of B. amyloliquefaciens CDD5 and B. amyloliquefaciens CPD4 reached 1.14 U/mL and 8.02 U/mL, respectively, at 48 hr. The proteases from the three B. amyloliquefaciens strains showed the highest activities within a pH range of 8.0-9.0 at $50^{\circ}C$, and casein was found to be the preferred substrate on evaluating enzyme activity in the substrate specificity assay. The B. amyloliquefaciens strains exhibited maximal growth when the nutrient broth medium had an initial pH within the range of 5.0-10.0, 6-9% sodium chloride (NaCl), and 5% glucose. B. amyloliquefaciens CDD5 exhibited a low ${\alpha}$-glucosidase inhibition rate (5.32%), whereas B. amyloliquefaciens CPD4 and B. amyloliquefaciens CGD3 exhibited relatively higher inhibition rates of 96.89% and 97.55%, respectively.


Supported by : 한국연구재단


  1. FAO/WHO (2002) Joint FAO/WHO (Food and Agriculture Organization/ World Healthb Organization) working group report on drafting guidelines for the evaluation of probiotics in food. London, Ontatiio, Canada
  2. Homma N (1998) Bifidobacteria as a resistance factor in human beings. Biosci Microflora, 7, 35-43
  3. Shin HJ, Bang JH, Choi HJ, Kim DW, Ahn CS, Jeong YK, Joo WH (2012) Probiotic potential of indigenous Bacillus sp. BCNU 9028 isolated from Meju. Korean J Food Sci Technol, 22, 605-612
  4. Hairul Islam VI, Prakash Babu N, Pandikumar P, Ignacimuthu S (2011) Isolation and characterization of putative probiotic bacterial strain, Bacillus amyloliquefaciens, from North East Himalayan soil based on in vitro and in vivo functional properties. Probiotics Antimicrob Proteins, 3, 175-185
  5. Gupta R, Beg QK, Khan S, Chauhan B (2002) An overview on fermentation, downstream processing and properties of microbial alkaline proteases. Appl Microbiol Biotechnol, 60, 381-395
  6. Bae YE, Yoon KH (2012) Production and characterization of thermostable protease from Bacillus licheniformis isolated from Korean traditional soybean paste. Korean J Microbiol, 48, 298-304
  7. Park CS, Min DK, Ahn YS, Lee JH, Hong SK, Kim JH, Kang DK (2002) Isolation and characteristics of soy protein-degrading strain, Bacillus subtilis EB464. J Microbiol Biotechnol, 30, 210-215
  8. Ahn YS, Kim YS, Shin DH (2006) Isolation, identification and fermentation characteristics of Bacillus sp. with high protease activity from traditional Cheonggukjang. Korean J Food Sci Technol, 38, 82-87
  9. Jung HK, Jeong YS, Youn KS, Kim DI, Hong JH (2009) Quality characteristics of soybean paste (Doejang) prepared with Bacillus subtilis DH3 expressing high protease levels and deep-sea water. Korean J Food Preserv, 16, 348-354
  10. Kim DY, Lee ET, Kim SD (2003) Purification and characterization of fibrinolytic enzyme produced by Bacillus subtilis K7 isolated from Korean traditional soy sauce. J Korean Soc Agric Biol Chem, 46, 176-182
  11. Yang SJ, Lee DH, Park HM, Jung HK, Park CS, Hong JH (2014) Amylase activity and characterization of Bacillus subtillis CBD2 isolated from Doenjang. Korean J Food Preserv, 21, 286-293
  12. Bell DS (2004) Type 2 diabets melitus : what is the optimal treatment regimen? Am J Med, 116, 23-29
  13. Lebovitiz HE (1997) Alpha-glucosidase inhibitors. Endocrinol Metab Clin North Am, 26, 539-551
  14. De Melo EB, da Silveira Gomes A, Cavalho I (2006) ${\alpha}$- and ${\beta}$-glucosidase inhibitors : chemical structure and biological activity. Tetrahedron 62, 10277-10302
  15. Courageot MP, Frenkiel MP, Dos Santos CD, Deubel V, Depres P (2000) Alpha-glucosidase inhibitors reduce dengue virus production by affecting the initial steps of virion morphogenesis in the endoplasmic reticulum. J Virol, 74, 564-572
  16. Kim SD (2014) Isolation and characterization of ${\alpha}$ -glucosidase inhibitor produced by Bacillus sp. SKU31-1 Strain. Korean J Microbiol, 50, 381-383
  17. Schmidit DD, Frommer w, Junge B, Muller L, Wingender W, Truscheit E, Schafer D (1977) Alpha-glucosidase inhibitors. Naturwissenschaften, 64, 535-536
  18. Zhu YP, Yin LJ, Cheng YQ, Yamaki K, Yutaka M, Su YC, Li LT (2008) Effect of source of carbon and nitrogen on production of ${\alpha}$-glucosidase inhibitor by a newly isolated srain of Bacillus subtillis B2. Food Chem, 109, 737-742
  19. Myoung KS, Heo G, Yoon SY, Shim JJ, Lee JH, Lim KS, Huh CS (2008) Inhibitory effect of Bacillus subtillis Y3-7 culture broth on ${\alpha}$-glucosidase activity. Korean J Food Sci Techol, 40, 558-561
  20. Kunitz M (1947) Crystalline soybean trypsin inhibitor. J Gen Physiol, 30, 291-295
  21. Watanabe J, Kawabata J, Kurihara H, Niki R (1997) Isolation and identification of alpha-glucosidase inhibitors from tochu-cha (Eucommia ulmoides). Biosci Biotech Biochem, 61, 177-178
  22. Jung HJ (2012) Preparation of functional deonjang using Bacillus amyloliquefacie -ns isolated from traditional commercial deonjang. MS Thesis. Kyung hee University. Korea. p 16
  23. Lee NR, Go TH, Lee SM, Hong CO, Park KM, Park GT, Hwang DY, Son HJ (2013) Characteristics of Chungkookjang prepared by Bacillus amyloliquefaciens with different soybeans and fermentation temperatures. J Microbiol, 49, 71-77
  24. Lim SI, Kim HK, Yoo JY (2000) Characteristics of protease produced by Bacillus subtillis PCA 20-3 isolated from Korean traditional Meju. Korean J Food Sci Techol, 32, 154-160
  25. Kim JY (2007) Isolation and characterization of an alkaline protease produced by Bacillus subtilis JK-1. J Microbiol, 43, 331-336
  26. Hwang SY (1995) Purification and characterization of extracellular serine protease produced from Bacillus sp. KUN-17. J Korean Soc Appl Biol Chem, 23, 53
  27. Moon SY, Oh TK, Rho HM (1994) Purification and characterization of an extra cellular alkaline protease from Bacillus subtilis RM 615. Korean Biochem J, 27, 323-329
  28. Kim TH, Park SH, Lee DS, Kwon TK, Kim JH, Hong SD (1990) Properties of alkaline protease produced by an alkalophilic Bacillus sp. J Microbiol Biotechnol, 18, 159-163
  29. Bang SH, Jeong IS (2011) Characterization of an alkaline protease from an alkalophilic Bacillus peseudofirmus HS-54. Korean J Microbiol, 47, 194-199
  30. Lim SI, Kwak EJ, Choi SY, Yoo JY (2002) Characteristics of protease produced by Rhizopus stolonifer, Rhizopus oryzae and Absidia corymbifera from Korean traditional Meju. J Korean Soc Food Sci Nutr, 31, 211-215
  31. Byun YG, Kim SH, Joo HK, Lee GS, Yim MH (1998) Purification and characterization of protease produced by Bacillus subtilis YG-95. J Korean Soc Appl Biol Chem, 41, 349-354
  32. Bae HC, Choi SH, Na SH, Nam MS (2012) Characteristics of ${\alpha}$-amylase and protease produced from Bacillus amyloliquefacies CNL-90 isolated from malt grain. J Anim Sci Technol, 54, 133-139
  33. Lee JY, Jeong SJ, Cho MJ, Cho KM, Kim GM, Shin JH, Lee KC, Kim JS, Kim JH (2014) Fibrinolytic activities of Bacillus species isolated from traditional fermented soyfoods. J Agric Life Sci, 48, 163-173
  34. Kim JW (2009) Screening of ${\alpha}$-glucosidase inhibitor producing Bacillus subtillis and its application. MS Thesis. Kangwon University, Korea, p 40-43
  35. Kim SH, Lee JY, Whang GY, Cho YS, Park YS, Kang KD, Seong SI (2011) Isolation and identification of a Bacillus sp. producing ${\alpha}$-glucosidase inhibitor 1-deoxynojirimycin. J Microbiol Biotechnol, 39, 49-55

Cited by

  1. Isolation and Characterization of Protease Producing B. amyloliquefaciens JH-35 from Food Waste vol.35, pp.4, 2016,