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A high-yielding lactic acid bacterium and its method for preparing calcium lactate by fermenting eggshells

A technology of lactic acid bacteria and calcium lactate, applied in the field of microorganisms, can solve the problems of increasing economic benefits, high energy consumption of the calcination method, environmental pollution, etc., and achieve the effects of overcoming environmental pollution, high purity, and high utilization rate of eggshells

Active Publication Date: 2015-10-14
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current research, the preparation of calcium lactate from eggshells mainly adopts chemical methods, and the most studied method is the calcination method. The calcination method consumes a lot of energy and costs high. Calcination produces a large amount of carbon dioxide and dust, which pollutes the environment.
The hydrothermal method and the direct neutralization method avoid the environmental pollution caused by high-temperature calcined eggshells, but these two methods require a large amount of lactic acid to be purchased, and the cost is relatively high
The preparation of calcium lactate by microbial method is an effective way to reduce pollution, increase economic benefits and reduce costs. At present, there is no literature report on the direct preparation of calcium lactate from eggshells by microbial fermentation.

Method used

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  • A high-yielding lactic acid bacterium and its method for preparing calcium lactate by fermenting eggshells
  • A high-yielding lactic acid bacterium and its method for preparing calcium lactate by fermenting eggshells
  • A high-yielding lactic acid bacterium and its method for preparing calcium lactate by fermenting eggshells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The isolation of embodiment 1 bacterial strain

[0037] The separation medium is MRS medium, purchased from Qingdao Haibo Biotechnology Co., Ltd., and the ingredients include: peptone 10.0g, beef powder 8.0g, yeast powder 4.0g, glucose 20.0g, sodium acetate 5.0g, dipotassium hydrogen phosphate 2.0g, diammonium hydrogen citrate 2.0g, Tween 801.0g, magnesium sulfate 0.2g, manganese sulfate 0.04g, agar 14g, distilled water 1000mL, pH value 6.5±0.2 (25℃). When using, weigh 66.2g of this product, dissolve it in 1000mL of distilled water by heating, autoclave at 121℃ for 15min, and set aside.

[0038] Lactic acid bacteria re-screening verification medium MC medium, purchased from Qingdao Haibo Biotechnology Company, contains: soybean peptone 5.0g, beef extract powder 3.0g, yeast extract powder 3.0g, glucose 20.0g, lactose 20.0g, calcium carbonate 10.0 g, agar 15.0g, neutral red 0.05g, distilled water 1000mL, pH 6.0 (25°C). When using, weigh 80.0g of this product, heat to bo...

Embodiment 2

[0045] Mutation and identification of embodiment 2 bacterial strains

[0046] 2.1 Study the effects of ultraviolet mutagenesis and nitrosoguanidine mutagenesis on the ability of lactic acid production:

[0047] 2.1.1 Ultraviolet (UV) mutagenesis

[0048] Take the slant of the bacteria, add 10mL of sterile water to make a suspension, take 0.1mL and spread it on 8 plates, culture it anaerobically at 37°C for 24h, and then place it under a 20W ultraviolet lamp for mutagenesis, and the irradiation distance is 30cm. Irradiate for 0.5min, 1.0min, 1.5min, 2.0min, 2.5min, 3min, 3.5min respectively, take the plate without UV irradiation as the control (CK), and then culture anaerobically at 37℃ for 24h. Spot the culture on the MC solid plate, culture anaerobically at 37°C for 42 hours, measure the diameter of the calcium-dissolving circle with a vernier caliper, measure 3 calcium-dissolving circles, take the average value, and compare the size of the calcium-dissolving circle at diffe...

Embodiment 3

[0094] The research of embodiment 3 strains producing lactic acid ability and fermentation time

[0095] 1. Acid production capacity of the strain

[0096] The components of the fermentation medium are configured as follows: peptone 0.5g, tryptone 0.5g, yeast extract 1.0g, glucose 10g, CaCl 2 0.008%, MgSO 4 0.192%, K 2 HPO 4 0.04%, KH 2 PO 4 0.04%, NaHCO 3 0.4%, NaCl0.08%, distilled water 100mL. Sterilize at 115°C for 20 minutes. Neutralizer calcium carbonate 6g, sterilized separately. Insert the logarithmic phase seed solution of lactic acid bacteria isolated in the present invention with an inoculation amount of 2%, and take Lactobacillus bulgaricus, which is the most commonly used strain for industrial fermentation to produce lactic acid at present, as a control. Incubate at 35°C for 72 hours on a 180r / min shaker. Place it at 85°C for 30 minutes, kill the bacteria, and filter. Take 1.0mL of the filtered fermentation broth, add 50mL of distilled water, then add 4....

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Abstract

The invention discloses a high-yield lactic acid bacterium which is Enterococcus mundtii HNMD-25, is collected in China general microbiological culture collection center (CGMCC), and has a collection number of CGMCC NO.8699. The invention further discloses application of the high-yield lactic acid bacterium to preparation of calcium lactate by fermenting calcium sources and a method for preparing calcium lactate by using eggshells. According to the method for preparing calcium lactate, provided by the invention, the eggshell utilization rate is high and reaches 77.98%, and about 2.5kg of calcium lactate pentahydrate can be prepared from 1kg of the eggshells; the glucose conversion rate is high, reaches 79.3% at most and is higher than that of common industrial strains; the calcium lactate purity is high and can be higher than 99% after fermentation products are refined.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a lactic acid bacterium with high lactic acid production and a method for preparing calcium lactate by fermenting eggshells. Background technique [0002] Enterococcus (Enterococcus) is a Gram-positive coccus widely distributed in nature and in humans and animals. Enterococcus mundtii (Enterococcus mundtii) is a type of enterococcus that is mostly reported as producing bacteriocin, which can kill or inhibit some pathogenic bacteria. Among them, Enterococcus mundtii ST4SA has been listed as a domestically preserved probiotic strain. Reports of Enterococcus monsonii producing antibiotics and antiviral peptides are very common. It has been reported that Enterococcus mundtii ST4SA and Lactobacillus plantarum423 Infant milk formula as a matrix as probiotics. Enterococcus has the ability to produce lactic acid, but there is no report on the production of lactic acid by Entero...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/56C12R1/46
Inventor 马美湖邓素枫金永国耿放
Owner HUAZHONG AGRI UNIV
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