Production of Nattokinase Using Bacillus natto NRRL 3666: Media Optimization, Scale Up, and Kinetic Modeling

다국어 초록 (Multilingual Abstract)

Fermentative production of nattokinase, a potent fibrinolytic enzyme using Bacillus natto NRRL 3666 was optimized by statistical optimization methods at shake flask. In addition, the production scheme based on optimum medium was scaled-up in 5-L lab-scale fermenter containing 2.5 L working volume. Further, unstructured mathematical models were proposed for kinetics of batch fermentation at shake flask as well as fermenter level for nattokinase. Enhanced production of nattokinase from 188±2.4 to 1,190.68±11 U/mL within 40 hr was achieved at shake flask. Nattokinase production improved to 1,932 U/mL in fermenter, which was 1.6 fold higher than shake flask. The production process was significantly reduced to 26 hr in the fermenter. The proposed models showed better prediction for experimental data with respect to biomass formation (R2>0.96), enzyme production (R2>0.99), and substrate utilization (R2>0.96). The present work showed successful optimization of medium for hyperproduction of nattokinase.

참고문헌 (Reference)

1 Fujita M, "Transport of nattokinase across the rat intestinal tract" 18 : 1194-1196, 1995

2 Fujita M, "Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat" 18 : 1387-1391, 1995

3 Urano T, "The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis cleaves and inactivates plasminogen activator inhibitor type-I" 276 : 24690-24696, 2001

4 Cheong-Il Ji, "The Processing Optimization of Caviar Analogs Encapsulated byCalcium-Alginate Gel Membranes" 한국식품과학회 16 (16): 557-564, 2007

5 Saran S, "Statistical optimization of conditions for protease production from Bacillus sp. and its scale-up in a bioreactor" 141 : 229-239, 2007

6 Nafise Shahbazian, "Statistical Optimization of the Medium Components for the Production of Protopectinases by Bacillus subtilis" 한국식품과학회 18 (18): 442-448, 2009

7 Shaileshkumar D. Sawale, "Statistical Optimization of Media for Dextran Production by Leuconostoc sp., Isolated from Fermented Idli Batter" 한국식품과학회 19 (19): 471-478, 2010

8 Kim W, "Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK11-4 screened from cheonggukjang" 62 : 2482-2488, 1996

9 Peng Y, "Purification and characterization of a fibrinolytic enzyme produced by Bacillus amyloliquefaciens DC-4 screened from douchi, a traditional chinese soybean food" 34 : 45-52, 2003

10 Chang CT, "Potent fibrinolytic enzyme from a mutant of Bacillus subtilis IMR-NK1" 48 : 3210-3216, 2000

11 Liu J, "Optimization of nutritional conditions for nattokinase production by Bacillus natto NLSSE using statistical experimental methods" 40 : 2757-2762, 2005

12 Jang-Hyun Lee, "Optimization of Medium Composition for Production of the Antioxidant Substances by Bacillus polyfermenticus SCD Using Response Surface Methodology" 한국식품과학회 18 (18): 959-964, 2009

13 Juliana Alves Macedo, "Optimization Studies for the Production of Microbial Transglutaminase from a Newly Isolated Strain of Streptomyces sp." 한국식품과학회 17 (17): 904-911, 2008

14 Chen P, "Medium optimization for the production of recombinant nattokinase by Bacillus subtilis using response surface methodology" 23 : 1327-1332, 2007

15 Mercier P, "Kinetics of lactic acid fermentation on glucose and corn by Lactobacillus amylophilus" 55 : 111-121, 1992

16 Daverey A, "Kinetics of growth and enhanced sophorolipids production by Candida bombicola using a low-cost fermentative medium" 160 : 2090-2101, 2010

17 Calik P, "Influence of pH conditions on metabolic regulations in serine alkaline protease production by Bacillus licheniformis" 31 : 685-697, 2002

18 Sumi H, "In vitro and in vivo fibrinolytic properties of nattokinase" 89 : 1267-, 1992

19 Sumi H, "Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase" 84 : 139-143, 1990

20 Chen PT, "Enhanced production of recombinant nattokinase in Bacillus subtilis by the elimination of limiting factors" 28 : 1595-1600, 2006

21 Revankar MS, "Enhanced production of laccase using a new isolate of white rot fungus WR-1" 41 : 581-588, 2006

22 Artemov AV, "Effect of polyelectrolytes on serine proteinase secretion by Bacillus subtilis" 262 : 33-35, 1990

23 Suzuki Y, "Dietary supplementation of fermented soybean, natto, suppresses intimal thickening, and modulates the lysis of mural thrombi after endothelial injury in rat femoral artery" 73 : 1289-1298, 2003

24 Fisher SH, "Control of carbon and nitrogen metabolism in Bacillus subtilis" 45 : 107-135, 1991

25 Benkortbi O, "Batch kinetics and modelling of Pleuromutilin production by Pleurotus mutili" 36 : 14-18, 2007

26 Agrebi R, "BSF1 fibrinolytic enzyme from a marine bacterium Bacillus subtilis A26: Purification, biochemical and molecular characterization" 44 : 1252-1259, 2009

27 Sumi H, "A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese natto: A typical and popular soybean food in the Japanese diet" 43 : 1110-1111, 1987

28 Omura K, "A newly derived protein from Bacillus subtilis natto with both antithrombotic and fibrinolytic effects" 99 : 247-251, 2005

29 Brooks JR, "A modified method for total carbohydrate analysis of glucose syrups, maltodextrins, and other starch hydrolysis products" 63 : 465-466, 1986

30 Liu JZ, "A mathematical model for gluconic acid fermentation by Aspergillus niger" 14 : 137-141, 2003