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Bacillus coagulans and its use in preparation of L-lactic acid

A technology of Bacillus coagulans and Bacillus coagulans, which is applied in the application field of Bacillus coagulans and its production of L-lactic acid by fermenting lignocellulose, which can solve problems such as application limitations and output to be improved, and reduce production costs and bacteria pollution The effect of low probability and high fermentation temperature

Pending Publication Date: 2017-10-03
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
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Problems solved by technology

[0004] For the above problems, the Chinese invention patent application publication number CN102690764 discloses a Bacillus coagulans used to prepare L-lactic acid and its application method. The strain can use five-carbon sugar or six-carbon sugar as raw material to obtain L-lactic acid, and its yield is also high, but this strain is mainly based on the by-products produced in the xylitol production process, especially the xylose extracted from the corn cob hydrolyzate, and it is produced during the fermentation process. need to add carbon source, so its application is greatly limited
[0005] The Chinese invention patent with the application publication number CN102643874 discloses a method for Bacillus to produce polymer-grade L-lactic acid using corn stalk hydrolyzate, and specifically discloses that the Bacillus can use corn stalk hydrolyzate as the only carbon source to ferment L-lactic acid, which The yield of L-lactic acid can reach 80g / l (80gL-lactic acid per liter of fermentation broth), although it can also use corn stalk hydrolyzate to produce L-lactic acid, but its yield needs to be improved

Method used

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  • Bacillus coagulans and its use in preparation of L-lactic acid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Isolation and screening of embodiment 1 bacterial strain

[0026] The culture medium used in the experiment process is as follows:

[0027] Enrichment medium: Each liter of medium contains 10g of xylose, 10g of yeast powder, and the rest is distilled water, pH 6.0, sterilized at 115°C for 20min.

[0028] Fermentation medium: Each liter of medium contains 50g of xylose, 10g of yeast powder, and the rest is distilled water, pH 6.0, sterilized at 115°C for 20min.

[0029] Growth medium: Each liter of medium contains 30g of xylose, 10g of yeast powder, CaCO 3 10g, 15-20g agar powder, pH 6.5, sterilized at 115°C for 20min.

[0030] Seed medium: Each liter of medium contains 50g of glucose, 10g of yeast powder, and CaCO 3 20g, the rest is distilled water, pH 6.5, sterilized at 115°C for 20min.

[0031] Treatment medium: gas-exploded corn stalk hydrolyzate 100-150ml, yeast powder 10g, CaCO 3 30g, pH6~7, sterilized at 115°C for 20min.

[0032] Collect compost samples fr...

Embodiment 2

[0034] Colony Morphological Characteristics and Physiological and Biochemical Characteristics of Example 2 Bacterial Strain BC-TJ

[0035] After the strain BC-TJ was inoculated on the growth medium and grown for 24 hours, it was observed that the colony was round, with a smooth surface, milky white, and smooth edges. Gram staining was positive, the cell shape was rod-shaped, the cell size was (0.7-0.9)×(3-5) μm, and endophytic spores. The suitable growth temperature of the strain is 30°C-60°C, and the optimum growth pH is 5.5-6.5. In physiological and biochemical tests, gelatin liquefaction, indole test and glucose fermentation gas production are negative; starch hydrolysis and V.P tests are positive, and can grow under both aerobic and anaerobic conditions at 60°C. See Table 1 for specific experimental results.

[0036] Table 1 Physiological and biochemical experiment results of strain BC-TJ

[0037]

[0038] Note: "+" means positive; "-" means negative.

Embodiment 3

[0039] 16S rDNA sequence analysis of Example 3 bacterial strain BC-TJ

[0040] In order to identify the isolated strain BC-TJ more accurately, the 16S rDNA sequence of the strain BC-TJ was amplified and compared. Firstly, the genomic DNA of the strain BC-TJ was extracted, which was used as a template for 16S rDNA sequence amplification.

[0041] The strain BC-TJ that was cultured overnight was collected by centrifugation at 12,000 rpm at room temperature for 1 min, and the bacterial DNA was extracted using a bacterial DNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.), according to the instructions.

[0042] 16S rDNA sequence amplification primers:

[0043] 27f: 5'-AGAGTTTGATCCTGGCTCAG-3'

[0044] 1492r: 5'-GGYTACCTTACGACTT-3'

[0045] Reaction conditions: pre-denaturation at 95°C for 5 min; denaturation at 5°C for 30 s, annealing at 54°C for 30 s, extension at 72°C for 1 min, a total of 35 cycles; and extension at 72°C for 10 min.

[0046] Reaction syst...

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Abstract

The invention provides Bacillus coagulans BC-TJ or its passage product Bacillus. The Bacillus coagulans BC-TJ has a preservation number of CGMCC NO. 14044. The Bacillus coagulans BC-TJ is a facultative anaerobic strain, can grow under aerobic and anaerobic conditions, has a high fermentation temperature, reduces contaminative microbe pollution probability during the fermentation process, is free of sterilization of the equipment, realizes open fermentation and reduces energy and the labor cost. The Bacillus coagulans BC-TJ utilizes lignocellulose hydrolyzate as a raw material to produce L-lactic acid, realizes a yield of 160+ / -1.5 g / l and optical purity of 100% or more and has a fermentation production capacity of 2.2 g / l / h. In the fermentation process, reductive sugar supply is avoided and a production cost is further reduced. The Bacillus coagulans BC-TJ has the great potential in large-scale industrial production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bacillus coagulans and its application to produce L-lactic acid by fermenting lignocellulose. Background technique [0002] Lactic acid is a versatile organic acid widely used in food, medicine, textile and chemical industries. As a multifunctional green platform compound, lactic acid is considered to be a promising fine chemical with good development potential. Traditional lactic acid bacteria production methods are mainly divided into chemical synthesis and microbial fermentation. Compared with the traditional chemical synthesis method, the microbial fermentation method can produce optically pure L-lactic acid or D-lactic acid according to different needs through different strains. [0003] At present, there are two bottlenecks in the production of lactic acid by microbial fermentation. On the one hand, the cost of using fermentable sugars such as glucose and starch in the fer...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/56C12R1/07
CPCC12N1/20C12P7/56C12N1/205C12R2001/07
Inventor 张东远赫荣琳
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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