阅读说明：本技术 一种提高表面活性素产量的方法 (Method for improving surfactant yield ) 是由 程景胜 唐敏慧 魏权石 元英进 于 2019-10-21 设计创作，主要内容包括：本发明公开了一种提高表面活性素产量的方法,包括如下步骤：将保藏编号为CGMCC No.7097的解淀粉芽孢杆菌(Bacillus amyloliquefaciens)HM618种子接入发酵培养基中,培养；发酵培养基包括：蔗糖,酵母粉,氨基酸,NaNO<Sub>3</Sub>,K<Sub>2</Sub>HPO<Sub>4</Sub>,KH<Sub>2</Sub>PO<Sub>4</Sub>,MgSO<Sub>4</Sub>·7H<Sub>2</Sub>O,FeCl<Sub>2</Sub>,ZnCl<Sub>2</Sub>,MnCl<Sub>2</Sub>·4H<Sub>2</Sub>O,pH自然,余量为水；所述氨基酸为丙氨酸、精氨酸、半胱氨酸、组氨酸、甘氨酸、赖氨酸、蛋氨酸、脯氨酸、丝氨酸、鸟氨酸和苏氨酸中至少一种。实验证明,本发明的方法,可以提高保藏编号为CGMCC 7097的解淀粉芽孢杆菌HM-618的生产表面活性素的产量。(The invention discloses a method for improving the yield of surfactin, which comprises the following steps: inoculating a Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HM618 seed with the preservation number of CGMCC No.7097 into a fermentation culture medium for culture; the fermentation medium comprises: sucrose, yeast powder, amino acid, NaNO 3 ，K 2 HPO 4 ，KH 2 PO 4 ，MgSO 4 ·7H 2 O，FeCl 2 ,ZnCl 2 ，MnCl 2 ·4H 2 O, natural pH and the balance of water; the amino acid is at least one of alanine, arginine, cysteine, histidine, glycine, lysine, methionine, proline, serine, ornithine and threonine. Experiments prove that the method can improve the growth of the bacillus amyloliquefaciens HM-618 with the preservation number of CGMCC 7097Yield of surfactant.)

1. A method for improving the yield of surfactin is characterized by comprising the following steps: the strain of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HM-618 with the preservation number of CGMCC No.7097 is selected according to the biomass OD₆₀₀Inoculating the strain into a fermentation medium at an inoculation density of 0.1-0.3, and culturing at 35-39 ℃ and a rotation speed of 100-300 rpm for 110-130 h; obtaining the surfactant.

The fermentation medium comprises the following components in g/L: 50-70 parts of cane sugar, 14-16 parts of yeast powder, 0.3-5 parts of amino acid and NaNO₃8～12，K₂HPO₄7～8，KH₂PO₄3～5，MgSO₄·7H₂O 2～4，FeCl₂0.01～0.03,ZnCl₂0.01～0.03，MnCl₂·4H₂0.01-0.03 percent of O, natural pH and the balance of water; the amino acid is at least one of alanine, arginine, cysteine, histidine, glycine, lysine, methionine, proline, serine, ornithine and threonine.

Technical Field

The invention belongs to the technical field of fermentation engineering, and relates to a method for improving the yield of surfactant.

Background

The surfactant is composed of four amino acids (valine, leucine, aspartic acid and glutamic acid) and fatty acid chain, has high surface activity, and has the characteristics of no toxicity or low toxicity, biodegradability and no environmental pollution compared with chemical surfactants. It has excellent antibacterial, antiviral, antitumor and anti-mycoplasma activities and great application potential.

At present, reports of improving the yield of the surfactant by exogenously adding non-constituent amino acids (other than valine, leucine, aspartic acid and glutamic acid) are not found.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a method for improving the yield of the surfactant.

The technical scheme of the invention is summarized as follows:

a method for increasing the yield of surfactin, comprising the steps of: the strain of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HM-618 with the preservation number of CGMCC No.7097 is selected according to the biomass OD₆₀₀Inoculating the strain into a fermentation medium at an inoculation density of 0.1-0.3, and culturing at 35-39 ℃ and a rotation speed of 100-300 rpm for 110-130 h; obtaining the surfactant.

The fermentation medium comprises the following components in g/L: 50-70 parts of cane sugar, 14-16 parts of yeast powder, 0.3-5 parts of amino acid and NaNO₃8～12，K₂HPO₄7～8，KH₂PO₄3～5，MgSO₄·7H₂O 2～4，FeCl₂0.01～0.03,ZnCl₂0.01～0.03，MnCl₂·4H₂0.01-0.03 percent of O, natural pH and the balance of water; the amino acid is at least one of alanine, arginine, cysteine, histidine, glycine, lysine, methionine, proline, serine, ornithine and threonine.

The invention has the advantages that:

experiments prove that the method can improve the yield of the surfactant produced by Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HM-618 with the preservation number of CGMCC No. 7097.

Drawings

FIG. 1 shows the effect of fermentation media containing histidine, ornithine and serine, respectively, on the production of surfactin in Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HM-618 with the collection number of CGMCC NO. 7097.

FIG. 2 is a graph showing the effect of fermentation media containing ornithine in different concentrations on the production of surfactin by Bacillus amyloliquefaciens (HM-618) with the collection number of CGMCC NO. 7097.

FIG. 3 is a graph showing the effect of fermentation media containing combinations of ornithine and serine, ornithine and threonine, and serine and threonine, respectively, on the production of surfactant from Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HM-618 with the accession number of CGMCC NO. 7097.

Detailed Description

The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) related by the invention is named as Bacillus amyloliquefaciens HM-618. Is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No. 7097. The preservation address is microbial research institute of China academy of sciences, No. 3, Xilu No. 1, Beijing, Chaoyang, and the preservation time is 2013, 1 month and 8 days, and the product survives. The preservation number of the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HM-618 is CGMCC No.7097, which is abbreviated as HM-618.

The HM-618 seeds of each example were obtained by culturing HM-618 in HM-618 seed medium.

HM-618 seed Medium (g/L): 60 parts of glucose, 10 parts of yeast powder, 10 parts of peptone, 5 parts of NaCl, 10 parts of beef extract, natural pH and the balance of water.