蚕豆纳豆发酵工艺优化及其酶学性质
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摘要
为开发新型蚕豆产品,提高蚕豆附加值研究以蚕豆为原料,通过单因素实验和正交试验优化出蚕豆纳豆的最佳工艺,并对发酵得到的纳豆激酶进行酶学性质研究。结果表明,最佳工艺条件为:蚕豆浸泡48 h,常压蒸煮20 min,接种量为6%,40℃下发酵72 h,此时,纳豆激酶活力值最高为(4860.13±470.02) U·g~(-1),感官评分为(95.67±0.71)分。酶学性质结果表明,该酶在25～40℃具有较好的热稳定性;在pH6～9范围内较为稳定,保持相对酶活89.05%以上; Mg~(2+)和Ca~(2+)对纳豆激酶活性具有促进作用,Cu~(2+)、Fe~(3+)抑制作用明显,Zn~(2+)无明显作用。本研究为蚕豆产品的开发及蚕豆纳豆的产业化生产提供参考。
In order to develop new broad bean products and increase the added value of broad bean,this study was based on broad bean as main material.The single-factor experiment and orthogonal experiment were used to optimize the best process of natto by broad bean and then the enzymatic properties of nattokinase were studied.The results showed that the optimal process conditions were as follows: The broad bean were soaked in 48 h and steamed under atmospheric pressure for 20 min,the inoculation of 6% and fermented at 40 ℃ for 72 h.Under these conditions,the enzyme activity reached (4860.13 ± 470.02) U·g~(-1),and the sensory score was (95.67 ± 0.71) points.Enzymatic properties analysis showed that the nattokinase had a better thermal stability at 25 ℃ to 40 ℃.Meanwhile,the nattokinase had good stability at a pH range from 6 to 9,and the relative enzyme activity was kept over 89.05%.The metal ions of Mg~(2+) and Ca~(2+) had an effect on promoting the activity of nattokinase,and the metal ions of Cu~(2+) and Fe~(3+) had an obvious effect on inhibiting nattokinase. However,the metal ions of Zn~(2+) had no obvious effect.This study could provide a reference for the development of broad bean products and the industrial production of broad bean natto.
In order to develop new broad bean products and increase the added value of broad bean,this study was based on broad bean as main material.The single-factor experiment and orthogonal experiment were used to optimize the best process of natto by broad bean and then the enzymatic properties of nattokinase were studied.The results showed that the optimal process conditions were as follows: The broad bean were soaked in 48 h and steamed under atmospheric pressure for 20 min,the inoculation of 6% and fermented at 40 ℃ for 72 h.Under these conditions,the enzyme activity reached (4860.13 ± 470.02) U·g~(-1),and the sensory score was (95.67 ± 0.71) points.Enzymatic properties analysis showed that the nattokinase had a better thermal stability at 25 ℃ to 40 ℃.Meanwhile,the nattokinase had good stability at a pH range from 6 to 9,and the relative enzyme activity was kept over 89.05%.The metal ions of Mg~(2+) and Ca~(2+) had an effect on promoting the activity of nattokinase,and the metal ions of Cu~(2+) and Fe~(3+) had an obvious effect on inhibiting nattokinase. However,the metal ions of Zn~(2+) had no obvious effect.This study could provide a reference for the development of broad bean products and the industrial production of broad bean natto.
引文
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[12]兰佳佳,杨希娟,王生君.蚕豆加工利用综述[J].青海农林科技,2017(4):46-49.
[13]张华龙,王明成,陈定国.蚕豆菜肥两用种植模式初探及发展对策[J].湖北农业科学,2017,56(19):3619-3620,3757.
[14]Siah S,Konczk I,Wood J A,et al.Effects of roasting on phenolic composition and in vitro antioxidant capacity of Australian grown gaba beans(Viciafaba L.)[J].Plant Foods for Human Nutrition,2014,69(1):85-91.
[15]李宏梁,赵倩楠.芸豆纳豆发酵工艺条件的研究[J].中国调味品,2014,39(3):46-49.
[16]Lin N N,Lee Y F,Chi Y J,et al.Bacillus subtilis-fermented red bean(red bean natto)reduces hyperlipidemia levels in hamsters fed an atherogenic diet[J].Journal of Food Biochemistry,2017,41(1):1-11.
[17]王琳,高辰哲,刘丹怡,等.响应面法优化红豆纳豆的发酵工艺[J].中国酿造,2018(1):190-194.
[18]Ma Y Y,Liu Q B,Yang H W,et al.Study on the improvement of natto-production process[J].Advance Journal of Food Science and Technology,2015,7(9):704-708.
[19]Shih M C,Yang K T,Su S Y,et al.Optimization process of roasted broken black soybean natto using response surface methodology[J].Journal of Food Processing and Preservation,2013,37(5):474-482.
[20]金爽,谭金燕,白秀云,等.固载纳豆菌发酵鹰嘴豆产品的感官评价[J].粮油食品科技,2016,24(4):86-89.
[21]金爽,谭金燕,白秀云,等.响应面法优化固载纳豆菌发酵鹰嘴豆工艺[J].食品与发酵科技,2016,52(3):28-32.
[22]张杰,葛武鹏,杨希娟,等.纳豆微胶囊的制备及其稳定性[J].食品工业科技,2017,38(22):157-162.
[23]陈应运,魏玉西,苏东海,等.纳豆菌液态发酵蛤蜊制备ACE抑制肽的工艺研究[J].核农学报,2016,30(11):2189-2195.
[24]张杰,葛武鹏,张静,等.高产纳豆激酶的枯草芽孢杆菌优选及发酵工艺条件优化[J].食品工业科技,2015,36(8):202-205,209.
[25]Astrup T,Muller S.The fibrin plate method for estimating fibrinolytic activity[J].Archives Biochemistry Biophysics,1995,40:346-351.
[26]Yang Y G.Study on the enzymatic properties of nattokinase[C]//5th International Conference on Information Engineering for Mechanics and Materials,2015.
[27]齐少卿,李晨,卢海强,等.保加利亚乳杆菌异源表达纳豆激酶的性质研究[J].中国食品学报,2017,17(4):51-57.
[2]Sumi H,Hamada H,Tsushima H.A novel fibrinolytic enzyme(nattokinase)in the vegetable cheese natto;atypical and popular soy-bean food in the Japanese diet[J].Experientia,1987,43(10):1110-1111.
[3]Fujita M,Hong K,Ito Y,et al.Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat[J].Biological and Pharmaceutical Bulletin,1995,18(10):1387-1391.
[4]平谷一,中西晃一朗,须见洋行.血栓溶解剂:日本,89180834[P].
[5]Essam K.Fibrinolytic bacterial enzymes with thrombolytic Activity[M].Springer Briefs in Microbiology,2012:35-43.
[6]王常苏,孙晓彤,余洁,等.纳豆激酶高活性菌株BN-05鉴定及发酵工艺优化[J].中国酿造,2014,33(1):91-95.
[7]Ero M P,Ng C M,Mihsilovski T,et al.A pilot study on the serum pharmacokinetics of nattokinase in humans following a single,oral,daily dose[J].Altern ative Ther apies Health Medicine,2013,19(3):16-19.
[8]马国兴,潘峰,王庆波,等.纳豆激酶分子结构与潜在应用价值分析[J].食品与生物技术学报,2016,35(4):342-349.
[9]张杰,葛武鹏,陈瑛,等.纳豆激酶高产菌株的选育及固态发酵技术[J].食品科学,2016,37(3):151-156.
[10]Chandrasekaran S D,Vaithilingam M,Shanker R,et al.Exploring the in vitro thrombolytic activity of nattokinase from a new strain Pseudomonas aeruginosa CMSS[J].Jundishapur Journal of Microbiology,2015,8(10):1-8.
[11]Wu C,Gao C M,Lv S Y,et al.Construction of polylysine dendrimer nanocomposites carrying nattokinase and their application in thrombolysis[J].Journal of Biomedical Materials Research Part A,2017,106(2):440-449.
[12]兰佳佳,杨希娟,王生君.蚕豆加工利用综述[J].青海农林科技,2017(4):46-49.
[13]张华龙,王明成,陈定国.蚕豆菜肥两用种植模式初探及发展对策[J].湖北农业科学,2017,56(19):3619-3620,3757.
[14]Siah S,Konczk I,Wood J A,et al.Effects of roasting on phenolic composition and in vitro antioxidant capacity of Australian grown gaba beans(Viciafaba L.)[J].Plant Foods for Human Nutrition,2014,69(1):85-91.
[15]李宏梁,赵倩楠.芸豆纳豆发酵工艺条件的研究[J].中国调味品,2014,39(3):46-49.
[16]Lin N N,Lee Y F,Chi Y J,et al.Bacillus subtilis-fermented red bean(red bean natto)reduces hyperlipidemia levels in hamsters fed an atherogenic diet[J].Journal of Food Biochemistry,2017,41(1):1-11.
[17]王琳,高辰哲,刘丹怡,等.响应面法优化红豆纳豆的发酵工艺[J].中国酿造,2018(1):190-194.
[18]Ma Y Y,Liu Q B,Yang H W,et al.Study on the improvement of natto-production process[J].Advance Journal of Food Science and Technology,2015,7(9):704-708.
[19]Shih M C,Yang K T,Su S Y,et al.Optimization process of roasted broken black soybean natto using response surface methodology[J].Journal of Food Processing and Preservation,2013,37(5):474-482.
[20]金爽,谭金燕,白秀云,等.固载纳豆菌发酵鹰嘴豆产品的感官评价[J].粮油食品科技,2016,24(4):86-89.
[21]金爽,谭金燕,白秀云,等.响应面法优化固载纳豆菌发酵鹰嘴豆工艺[J].食品与发酵科技,2016,52(3):28-32.
[22]张杰,葛武鹏,杨希娟,等.纳豆微胶囊的制备及其稳定性[J].食品工业科技,2017,38(22):157-162.
[23]陈应运,魏玉西,苏东海,等.纳豆菌液态发酵蛤蜊制备ACE抑制肽的工艺研究[J].核农学报,2016,30(11):2189-2195.
[24]张杰,葛武鹏,张静,等.高产纳豆激酶的枯草芽孢杆菌优选及发酵工艺条件优化[J].食品工业科技,2015,36(8):202-205,209.
[25]Astrup T,Muller S.The fibrin plate method for estimating fibrinolytic activity[J].Archives Biochemistry Biophysics,1995,40:346-351.
[26]Yang Y G.Study on the enzymatic properties of nattokinase[C]//5th International Conference on Information Engineering for Mechanics and Materials,2015.
[27]齐少卿,李晨,卢海强,等.保加利亚乳杆菌异源表达纳豆激酶的性质研究[J].中国食品学报,2017,17(4):51-57.