阅读说明：本技术 利用混菌发酵提高表面活性素产量的方法 (Method for improving surfactant yield by mixed fermentation ) 是由 程景胜 唐敏慧 元英进 于 2019-10-21 设计创作，主要内容包括：本发明公开了一种利用混菌发酵提高表面活性素产量的方法,包括如下步骤：将解淀粉芽孢杆菌Bacillus amyloliquefaciens HM-618,CGMCC No.7097的种子和枯草芽孢杆菌Bacillus subtilis,CGMCC1.12939的种子分别按生物量OD<Sub>600</Sub>为0.1～0.3的接种密度接入发酵培养基中,在35～39℃、转速100～300rpm条件下培养110～130h,得到表面活性素；利用解淀粉芽孢杆菌Bacillus amyloliquefaciens HM-618,CGMCC No.7097和枯草芽孢杆菌Bacillus subtilis,CGMCC1.12939共培养能提高表面活性素的产量。(The invention discloses a method for improving the yield of surfactant by mixed fermentation, which comprises the following steps: respectively using seeds of Bacillus amyloliquefaciens HM-618 and CGMCC No.7097 and seeds of Bacillus subtilis and CGMCC1.12939 according to biomass OD 600 Inoculating 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 to obtain surfactant; the co-culture of Bacillus amyloliquefaciens HM-618 (CGMCC No. 7097) and Bacillus subtilis (CGMCC 1.12939) can improve the yield of the surfactin.)

1. A method for improving the yield of surfactin by mixed fermentation is characterized by comprising the following steps: respectively using seeds of Bacillus amyloliquefaciens HM-618 and CGMCC No.7097 and seeds of Bacillus subtilis and CGMCC1.12939 according to 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 to obtain surfactant;

the fermentation medium comprises 50-70 g/L of glucose, 4-6 g/L of yeast powder and NaNO₃8～12g/L，K₂HPO₄7～8g/L,KH₂PO₄3～5g/L,MgSO₄·7H₂O 2～4g/L,FeCl₂0.01～0.03g/L,ZnCl₂0.01～0.03g/L,MnCl₂·4H₂0.01-0.03 g/L of O, the balance of water and natural pH.

Technical Field

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

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 also has good antibacterial, antiviral, antitumor and anti-mycoplasma activities, and has great application potential.

The wild strain discovered at present almost has milligram-level surfactant yield, and the production, separation and purification costs are extremely high.

The co-culture fermentation can improve the yield of microbial products and obtain products with higher quality, but the research on improving the yield of the surfactant by co-culture microorganisms is not reported.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a method for improving the yield of surfactin by mixed fermentation.

A method for improving the yield of surfactin by mixed fermentation comprises the following steps: respectively using seeds of Bacillus amyloliquefaciens HM-618 and CGMCC No.7097 and seeds of Bacillus subtilis and CGMCC1.12939 according to 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 to obtain surfactant;

the fermentation medium comprises 50-70 g/L of glucose, 4-6 g/L of yeast powder and NaNO₃8～12g/L，K₂HPO₄7～8g/L,KH₂PO₄3～5g/L,MgSO₄·7H₂O 2～4g/L,FeCl₂0.01～0.03g/L,ZnCl₂0.01～0.03g/L,MnCl₂·4H₂0.01-0.03 g/L of O, the balance of water and natural pH.

THE ADVANTAGES OF THE PRESENT INVENTION

The co-culture of Bacillus amyloliquefaciens HM-618 (CGMCC No. 7097) and Bacillus subtilis (CGMCC 1.12939) can improve the yield of the surfactin.

Detailed Description

The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is named as Bacillus amyloliquefaciens HM-618 and is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms 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.

In the following examples and comparative examples, the preservation number of HM-618 of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is CGMCC No.7097 and abbreviated as HM-618.

Bacillus subtilis, CGMCC1.12939, is purchased from China general microbiological culture Collection center (CGMCC) in 2016 (11 months) in China, and is hereinafter referred to as Bacillus subtilis CGMCC 1.12939.

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.

The Bacillus subtilis CGMCC1.12939 seed of each embodiment is obtained by culturing Bacillus subtilis CGMCC1.12939 in a Bacillus subtilis CGMCC1.12939 seed culture medium.

Bacillus subtilis CGMCC1.12939 seed culture medium (LB culture medium) (g/L): tryptone 10, Yeast extract 5, NaCl 10)

The present invention will be further described with reference to the following examples.