纳豆芽孢杆菌Bacillus natto NK4液态发酵产纳豆激酶的工艺优化

为提高纳豆激酶的产量,采用单因素试验及正交试验对纳豆芽孢杆菌Bacillus natto NK4进行发酵条件优化,获得该菌种液体发酵产酶的最优条件。结果表明,纳豆芽孢杆菌产纳豆激酶最佳的发酵条件为接种量2%,培养基初始pH值6.0,培养温度34℃,发酵时间72 h。此时,纳豆激酶酶活力由848.28 IU/mL提高到1569.31 IU/mL。

Condition Optimization of Lipopeptide Production by Bacillus natto NT-6 in Solid-State Fermentation with Box-Behnken Design

Solid-state fermentation has certain advantages in improving the yield of lipopetide, Box-Behnken Design（BBD） was adopted to optimize the producing condition of the antibacterial lipopetide produced by Bacillus natto in this article. The optimal solid state fermentation conditions were obtained： 10 g solid medium（7 g of wheat bran, 3 g of soybean meal） with appropriate inorganic salt（glucose 0.67%,sodium glutamate 0.64%,（NH4）2SO40.15%, K2HPO40.10%）; moisture content 123.78%; inoculation amount 10%; cultivation temperature 36.75 ℃ and cultivation time 72.4 h. The maximum production of lipopetide is 61.76 mg/gds under such conditions. This is the first report on the optimization of lipopeptide fermentation conditions in solid-state fermentation by wheat bran and soybean meal with Bacillus natto NT-6 strain, and will contribute to the development of lipopetide production.

Construction and Expression of Methionine-rich and Lysine-rich Fusion Gene in Bacillus natto

Methionine and lysine are restrictive essential amino acids of livestock, they are also the most attentive indexes in the feed production to carry out the quality control and quality evaluation. Their contents in feed directly affect livestock protein synthesis. Bacillus natto has excellent probiotic properties. In this experiment, we used the genetic engineering method, fusion PCR technique, to connect methionine-rich gene （zein） from maize endosperm protein with lysine-rich gene （Cflr） from the pepper anther, then the fusion gene was inserted into the expression vector pHT43, and the recombinant plasmid pHT43/zein-Cflr was constructed. The recombinant plasmid was transferred into Bacillus natto, and induced by IPTG for the expression of the fusion gene. We found an apparent band at 40 ku site for the recombinant strain by SDS-PAGE. The contents of methionine and lysine were individually detected with HPLC, the quantities of methionine and lysine in the recombinant strain increased by 18.37% and 24.68% than the wild one, respectively. We also verified the stability of the recombinant bacterium during passaging, and found the stability was 100%. This study provided research-basis for the application of the recombined Bacillus natto as feed additive.

Optimization of Solid State Fermentation Conditions Using a Mixture of Bean Curd Residue and Marc with Bacillus natto

[Objective] The aim was to optimize the appropriate solid state fermentation(SSF)conditions.[Method] The optimization of solid state fermentation using a mixture substrate of bean curd residue and the marc with Bacillus natto was developed.[Result] The best fermentation condition optimized by the test of single factor and the orthogonal design respectively was mixing ratio of bean curd residue to marc 2∶1,substrate pH value 6,fermentation temperature 39 ℃,inoculum volume 10% and fermentation time 48 h.Under this optimized fermentation condition,the content of crude fiber in the substrate decreased from 107.8 mg/g before SSF to 56.2 mg/g after SSF,and the degeneration rate of crude fiber was 47.87%.[Conclusion] The bean curd residue in its palatability was enormously improved by SSF with Bacillus natto strain,which could be expected to be widely used as raw material of health foodstuff.

Study on the Antioxidant Component of Fermented Rice Bran by Bacillus Natto

Each component of rice bran fermentation(RBF)of Bacillus natto was extracted and its antioxidant activity was tested.By dint of thin layer chromatography,the antioxidant components were separated.The sample isolated from TLC for ethyl acetate phase and ether phase was analyzed by GC-MS.Results indicated that the antioxidant activity of large components in rice bran fermentation was low.The constituents of extracted phase had antioxidant activity apart from the petroleum ether phase.The constituents of ethyl acetate phase and Ether phase had best antioxidant activity,163 and 168 kU/g,respectively.The main composition of ethyl acetate phase was allyl group methyl-thioether and the main composition of ether phase included 3,5-dimethyl phenol,2,4-dipl-tert-Butyl and Fluazifop.

Study on Lysine and Methionine Content Promotion of Soybean Meal by Probiotic Fermentation Process

Soybean meal （SBM） is commonly used for livestock feeds, but its application in diets for livestock is limited due to some antinutritional factors. The contents of methionine and lysine of soybean meal were promoted by Bacillus natto and Leuconostoc mesenteroides fermentation, benefial for the livestock feeds. It was crude protein （CP） 56.8%, methionine 43.56 mg · g^-1, and lysine 74.87 mg · g^-1, cows fed a diet with FSBM milk yield raised 14.2%, the change in the milk protein, the lactose and the dry matter content had also obvious increase. This convenient technique offers helpful exploration for industrialization of soybean meal fermentation.