CN108977396B - Lactobacillus plantarum for high yield of lactic acid and application thereof - Google Patents
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Abstract
The invention discloses lactobacillus plantarum for high yield of lactic acid and application thereof, and belongs to the field of microbiology. The lactobacillus plantarum CGMCC No.16112 applied in the invention has large acid production amount and high acid production rate, can still keep good growth state under the condition of low pH value in the later fermentation period, has active enzyme system and accumulates high-content lactic acid; furthermore, the utilization rate of the lactobacillus plantarum on sucrose reaches 96.03%, and various sugars can be utilized; further, lactobacillus plantarum used in the present invention is a safe strain for food use; no other chemical substances are added, so that the health-care food is safe and healthy; furthermore, the lactobacillus plantarum can effectively improve the content of lactic acid substances in the fermented product and has a remarkable improvement effect on the texture characteristics of the product.
Description
Technical Field
The invention relates to lactobacillus plantarum for high yield of lactic acid and application thereof, belonging to the field of microbiology.
Background
The sour pulp is a coagulant widely used in the bean product industry, is generally fermented by microorganisms, and has the main effective component of organic acid, wherein lactic acid is the main contributor of the organic acid.
The existing production modes of lactic acid mainly comprise a chemical synthesis method, an enzyme conversion method and a microbial fermentation method, wherein the lactic acid produced by the microbial fermentation method is L-type, has high optical purity, no toxic or side effect on a human body, soft acidity and stable properties, and is suitable for an acidity regulator, a preservative, a pH regulator and the like (Liujin plum, the breeding of high-yield lactate acid radical mould strains in traditional distiller's yeast and the optimization of fermentation conditions [ D ] of Jiangxi agriculture university, 2015 ]).
Microorganisms commonly used for the production of lactic Acid include lactic Acid bacteria and Rhizopus strains, which were found by the Yang Guo et al (Yang Guo, Qiaojuan Yan, Zhengqiang Jiang, et al. effective production of lactic Acid from lactic Acid and carbohydrate by a new microorganism Biotechnol 2010,37:1137-1143.) DE GY18, which was fermented at 35 ℃ for 24h in sucrose solution capable of 20 g/L, producing 3 g/L left hand type lactic Acid Sachin 0-type lactic Acid Sachin R et al (Kadam S R, Patil SS, Bastawde K B, et al. strain of 5 active bacterium of 5 bacterium, Saccharum japonica L.R. (Kadam S R, Patil S, Bastawde K B, et al. S. strain K65, strain K12, strain of lactic Acid from Lactobacillus strain, K12, strain K12, strain K7-12, strain K7, strain K35, strain K7-12, strain K7, strain K35, strain K35, strain K7, strain K7, strain K35, strain.
In summary, in the lactic acid-producing strains, the yield of lactic acid is not high when the wild strain and the mutant strain utilize sugars such as sucrose as carbon sources, and the soybean product contains abundant sucrose, and meanwhile, lactobacillus plantarum is a commonly used lactic acid bacterium in fermented products, so research and development of lactobacillus plantarum capable of utilizing sucrose to produce lactic acid at high yield are expected to be used as a leavening agent for soybean product fermentation to improve the gel rate and texture of the soybean product, and the technical problem to be solved in the field is urgently needed.
Disclosure of Invention
In order to solve the existing problems, the invention provides lactobacillus plantarum with high lactic acid yield and application thereof; the technical scheme is as follows:
the invention aims to provide lactobacillus plantarum (L actinobacillus plantarum) with high lactic acid yield, which is preserved in China general microbiological culture Collection center (CGMCC) at 16.07.2018 with the preservation number of CGMCC No.16112 and the preservation address of Naja Kogyo No.1 Hopkins, Naja Kogyo No. 3, China academy of sciences microbial research institute.
The second purpose of the invention is to provide a microbial inoculum containing the lactobacillus plantarum.
The third purpose of the invention is to provide the application of the lactobacillus plantarum in the preparation of lactic acid.
The fourth purpose of the invention is to provide an application of the lactobacillus plantarum in preparation of bean products.
Optionally, the application of lactobacillus plantarum in the preparation of bean products comprises the application of lactobacillus plantarum in the preparation of sour soybean milk and bean curd.
Optionally, the application of lactobacillus plantarum in preparing the sour soybean milk comprises the following steps: taking fresh aseptic soybean milk as a raw material, adding yellow serofluid fermented by lactobacillus plantarum for 24h, preserving heat at 37 ℃ for 6h, adding other ingredients, homogenizing and packaging to obtain the sour soybean milk product.
Optionally, the application of lactobacillus plantarum in preparing bean curd comprises the following steps: washing activated Lactobacillus plantarum, and adjusting cell concentration of Lactobacillus plantarum to 109More than CFU/m L, freeze drying to obtain direct vat set starter, inoculating 1g starter powder into 1000m L soybean milk, fermenting at 37 deg.C for 4 hr, and adding other adjuvants to obtain sour bean curd product.
The fifth purpose of the invention is to provide the application of the lactobacillus plantarum in the preparation of pickled vegetables.
The sixth purpose of the invention is to provide an application of the lactobacillus plantarum in yogurt making.
The seventh purpose of the invention is to provide the application of the microbial inoculum containing the lactobacillus plantarum in the preparation of lactic acid.
The eighth purpose of the invention is to provide an application of the microbial inoculum containing the lactobacillus plantarum in the preparation of bean products.
Optionally, the soy product comprises a sour soy milk neutralized soy curd.
The ninth purpose of the invention is to provide the application of the microbial inoculum containing the lactobacillus plantarum in the preparation of pickled vegetables.
The tenth purpose of the invention is to provide the application of the microbial inoculum containing the lactobacillus plantarum in the preparation of the yoghourt.
The invention has the beneficial effects that:
the lactobacillus plantarum CGMCC No.16112 applied in the invention has the lactic acid yield of 8.02 g/L in 24h, which is higher than the lactic acid yield of naturally fermented yellow serofluid C1 and lactobacillus paracasei C2, the lactobacillus plantarum CGMCC No.16112 has the lactic acid output rate of 0.15 g/L/h and higher than 0.12 g/L/h of C1 and 0.08 g/L/h of C2 in 12h to 24h, which shows that the lactobacillus plantarum CGMCC No.16112 can still keep better growth and activate enzyme system and accumulate high content lactic acid under the condition of lower pH value in the later fermentation period, further, the utilization rate of the lactobacillus plantarum CGMCC No.16112 on cane sugar reaches 1615 percent and can utilize various sugars, further, the lactobacillus plantarum CGMCC No.16112 can produce 6.37 g/L in 6h and is a product for naturally preserving lactobacillus plantarum CGMCC No. 6 and lactobacillus paracasei C4835, the lactobacillus plantarum CGMCC No.16112 has the effect of improving the lactic acid yield of milk curd materials, and milk curd quality, and the milk products for improving the milk quality of milk products, wherein the lactobacillus plantarum CGMCC No.16112 has the effective and the safe fermentation rate of the lactobacillus plantarum CGMCC No.1, the safe fermentation of the lactobacillus plantarum CGMCC No.1, the lactobacillus plantarum CGMCC No. of the lactobacillus plantarum CGMCC No.16112 added in the lactobacillus plantarum CGMCC NO:
the lactobacillus plantarum CGMCC No.16112 is preserved in China general microbiological culture Collection center in 2018, 07, 16 and 16 months, and the preservation number is as follows: CGMCC No.16112, preservation Address: xilu No.1, Beijing, Chaoyang, Beijing, and institute for microbiology, China academy of sciences.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1a is a colony morphology diagram of Lactobacillus plantarum CGMCC No.16112 on a solid MRS medium;
FIG. 1b is a cytomicroscopic image of Lactobacillus plantarum CGMCC No. 16112;
FIG. 2 shows the growth of Lactobacillus plantarum CGMCC No. 1611224h;
FIG. 3 shows the acid production of yellow serofluid fermented in Lactobacillus plantarum CGMCC No. 1611224h;
FIG. 4 shows the sugar utilization of fermented yellow serofluid in Lactobacillus plantarum CGMCC No. 1611224h.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in detail with reference to the accompanying drawings.
The first embodiment is as follows: strain separation and identification method of lactobacillus plantarum
(1) Obtaining appropriate dilution gradient and culturing
0.5ml of bean curd sour milk sample is extracted from a bean curd sour milk sample of sheep spring fermented bean curd company of mugding county, Chuxiong province, Yunnan province, is added into sterile physiological saline water with the concentration of 4.5m L, 0.5m L bacterial liquid is repeatedly extracted and diluted into sterile physiological saline water with the concentration of 4.5m L, and the like, so that the concentration of the sample is diluted to 10 degrees in a gradient manner-6CFU/m L, taking 6 dilutions of 10-106Each bacterial suspension of CFU/m L is coated on MRS solid medium at 100 mu L, and is cultured for 24-48h at 37 ℃ in an inverted mode.
(2) Separating and purifying
Typical single colonies were picked, purified and isolated by plate three-zone streaking, and this culture selection procedure was repeated until all single colonies were purified.
(3) Gram stain and Catalase assay
Picking a single colony on a glass slide, performing smear, drying, fixing, primary dyeing, washing, mordanting, washing, decoloring, counterdyeing, washing, drying and microscopic examination, and recording a gram staining result; and picking single colonies on a glass slide, adding a 3% hydrogen peroxide solution, observing the generation of bubbles, recording the contact result of catalase, and reserving gram-positive colonies and catalase-negative colonies.
(4) Strain preservation
Selecting and purifying the single colony in a 5m L liquid MRS culture medium, standing and culturing at 37 ℃ for 20-24h, sucking 1m L bacterial liquid into a bacteria-preserving tube, centrifuging at 4000rpm for 5min, removing supernatant, adding 1m L30% sterile glycerol solution, resuspending, and preserving at-80 ℃.
(5) PCR amplification of 16S rDNA
Sucking liquid culture medium with 1m L, shaking and mixing uniformly, centrifuging, discarding supernatant, blowing and washing with 1m L sterile water, centrifuging, discarding supernatant, and using the supernatant as a template for colony PCR.
1) PCR system 25. mu. L, where Mix was 12.5. mu. L, 27F was 0.5. mu. L, 1492R was 0.5. mu. L, and ddH2O was 11.5. mu. L, the primers used were 27F: AGA GTT TGA TCC TGG CCT CA 20(SEQ ID NO:2) and 1492R: GGT TAC CTTGTT ACG ACT T19 (SEQ ID NO:3), the nucleotide sequence of the amplified fragment of the target amplified fragment length 1500 bp. is shown in SEQ ID NO:1, and the length is 1435 bp.
2) PCR conditions were as follows:
pre-denaturation temperature: 105 ℃ C
First-step denaturation: 7min at 95 ℃; 30s at 95 DEG C
And a second step of annealing: 30s at 55 DEG C
And a third step of extension: 90s at 72 DEG C
Cycle number: 30s at 95 ℃ for 33 cycles
The fourth step is finally extended: 5min at 72 DEG C
The fifth step is that: 10min at 12 DEG C
(6) Agar gel electrophoresis (80m L)
Weighing 0.8g of agarose, adding the agarose into a conical flask, adding 80m L1 xTAE, heating for 4min in a microwave discontinuous manner until the liquid is clear and transparent, slightly cooling, adding 2 per mill of nucleic acid dye, shaking uniformly without bubbles, pouring the mixture into a rubber plate groove, cooling for 40min, solidifying, placing the mixture into an electrophoresis groove, exhausting bubbles, sequentially adding PCR amplification products, adding 4 mu L PCR amplification products into each hole, taking out after 120V 30min gel running, placing the mixture into a gel electrophoresis imager for photographing and storing, recording the serial number of samples with successful PCR, and placing the successful PCR products into a refrigerator at-20 ℃ for storage.
(7)16S rDNA sequence analysis and identification
And (3) sending the sample with successful PCR to Huada gene for detection, carrying out B L AST retrieval by combining an NCBI strain sequence database according to a sequence result fed back by the Huada gene, and selecting strain information with the highest matching degree for result recording, wherein the result shows that the strain provided by the invention is lactobacillus plantarum (L actobacillus plantarum).
The bacterial colony characteristics of the strain are as follows: the form of the strain on a solid MRS culture medium is shown in figure 1a, and the colony is white and glossy after about 30 hours, and the edge of the colony is circular, opaque, convex and small. As shown in FIG. 1b, the microbial cells of Lactobacillus plantarum CGMCC No.16112 observed under an oil lens are short rod-shaped, and the gram-positive bacteria are purple in gram stain.
The name and source of the sample used in the example of the invention are shown in table 1, wherein the natural fermentation physalis alkekengi simulation liquid is prepared by inoculating 10% of collected bean curd physalis alkekengi into yellow serofluid and fermenting at 37 ℃ for 6 h.
The numbers, strain names and sources of lactic acid bacteria used in the examples of the present invention are shown in table 2, in which C1 represents a natural mixed strain.
TABLE 1 sample name and Source used in the invention
TABLE 2 lactic acid bacteria numbering, strain name and source used in the invention
Example two: growth curve of lactobacillus plantarum in MRS within 24h
Inoculating a strain preserved at the temperature of minus 80 ℃ into a 5m L liquid MRS culture medium according to the inoculation amount of 2%, culturing for 16-18h at the temperature of 37 ℃, activating for third generation culture, inoculating a bacterial liquid into the MRS liquid culture medium according to the inoculation amount of 2% for culturing for 24h, taking the bacterial liquid 1m L every 2h from 0h, shaking and uniformly mixing, measuring OD600 absorbance by using an ultraviolet spectrophotometer, paralleling for three times, and drawing a growth curve according to time, wherein the result is shown in figure 1, and the lactobacillus plantarum CGMCC No.16112 enters a stable period in 12 h.
Example three: pH change of Lactobacillus plantarum in yellow serofluid within 24h
Inoculating the strain preserved at-80 ℃ into a 5m L liquid MRS culture medium according to the inoculation amount of 2%, culturing for 16-18h at 37 ℃, activating for three generations, inoculating the bacterial liquid into a sterile yellow serofluid liquid culture medium according to the inoculation amount of 3%, culturing for 24h, taking the fermented yellow serofluid bacterial liquid every 2h from 0h, shaking and uniformly mixing, measuring the pH value by using a pH meter, paralleling for three times and recording, wherein the result is shown in figure 2.
As can be seen from FIG. 2, the pH value of Lactobacillus plantarum CGMCC No.16112 can be reduced from 6.25 to 3.86 in yellow serofluid, and the reduction degree of the pH value is stronger than that of the natural fermentation group C1 and the paracasei fermentation group C2.
Example four: lactic acid production capacity of lactobacillus plantarum in yellow serofluid within 24h
Inoculating a strain preserved at the temperature of minus 80 ℃ into a 5m L liquid MRS culture medium according to the inoculation amount of 2 percent, culturing for 24h at the temperature of 37 ℃, taking 1m L bacterial liquid when activating to the third generation, centrifuging for 5min at 4000rpm/min to obtain bacterial sludge, washing with sterile physiological saline for 2 times, then suspending in 1m L sterile physiological saline, inoculating into sterile yellow serous fluid according to the inoculation amount of 3 percent, culturing for 24h at the temperature of 37 ℃, sampling from 0h, taking 2m L fermented yellow serous fluid samples each time, centrifuging for 10min at 10000rpm, taking supernatant, filtering with a 0.22 mu m filter membrane, determining the organic acid substance composition, and performing three-time determination.
The chromatographic conditions of the instrument Agilent 1200, the chromatographic column of Diamonsil C18 (4.6mm × 250mm, 5 μm), the mobile phase of methanol, water and phosphoric acid (5: 95: 0.05, volume ratio), the flow rate of 0.6m L/min, the column temperature of 30 ℃, the detector of an ultraviolet detector, the detection wavelength of 210nm, the sample injection amount of 5 μ L and the elution time of 30 min.
The results of the lactic acid content of the strain used in the embodiment of the invention in fermenting yellow serofluid in 24h are shown in Table 3, and from Table 3, it can be known that in 12h, the lactic acid yield of Lactobacillus plantarum CGMCC No.16112 is 6.19 g/L, lower than the lactic acid yield of C1 of a naturally fermented yellow serofluid group, higher than the lactic acid yield of C2 of a Lactobacillus paracasei group, and stronger in the lactic acid production capacity of Lactobacillus paracasei, in 24h, the lactic acid yield of Lactobacillus plantarum CGMCC No.16112 is 8.02 g/L, higher than the lactic acid yields of C1 and C2 of a naturally fermented yellow serofluid group, and in 12h to 24h, the lactic acid production rate of Lactobacillus plantarum CGMCC No.16112 is 0.15 g/L/h, higher than 0.12 g/L/h of a C1 group and 0.08 g/L/h of a C2 group, and that in the later stage, the lactic acid content of the Lactobacillus plantarum CGMCC No.16112 can still keep the active strain in the growth state and the lactic acid content of the lactobacillus paracasei is higher than the CGMCC No. C1 group.
TABLE 3 lactic acid production of fermented yellow serofluid at different time points
Example five: sugar utilization capacity of lactobacillus plantarum in yellow serofluid within 24h
Inoculating a strain preserved at the temperature of minus 80 ℃ into a 5m L liquid MRS culture medium according to the inoculation amount of 2 percent, culturing for 24h at the temperature of 37 ℃, taking 1m L bacterial liquid when the strain is activated to the third generation, centrifuging for 5min at 4000rpm/min to obtain bacterial sludge, washing with sterile physiological saline for 2 times, then suspending in 1m L sterile physiological saline, inoculating into sterile yellow serofluid according to the inoculation amount of 3 percent, culturing for 24h at the temperature of 37 ℃, taking 2m L fermented yellow sauce water samples at two measurement points of 0h and 24h respectively, centrifuging for 10min at 10000rpm, taking supernatant, filtering with a 0.22 mu m filter membrane, measuring the sugar composition, and performing three times of measurement in parallel.
The chromatographic conditions are that an instrument Waters 600 is adopted, a chromatographic column is a Sugar-pakl C18 column (6.5mm × 300mm, 5 mu m), the mobile phase is ultrapure water, the flow rate is 0.4m L/min, the column temperature is 85 ℃, a detector is a differential refraction detector, the sample introduction amount is 10 mu L, and the elution time is 30 min.
The results of the change of the sugar content of the yellow serofluid fermented by the strain used in the embodiment of the invention in 24h are shown in figure 3, and as can be seen from figure 3, 4 of 5 sugars can be utilized by both the natural fermentation group C1 and the lactobacillus paracasei fermentation group C2, 5 sugars can be utilized by lactobacillus plantarum CGMCC No.16112, wherein the sucrose content can be reduced from 5.01 g/L to 0.20 g/L, the utilization rate can reach 96.03%, and the formation of organic acid is promoted.
Example six: acid production capacity of lactobacillus plantarum in soybean milk within 6h
Selecting high-quality soybeans with plump seed grains, washing the soybeans cleanly by using deionized water, soaking the soybeans for 10-12 hours at room temperature by using a soybean water ratio of 1/3.5 to enable the soybean grains to fully absorb water until the inner sides of soybean petals are basically flat, then draining the soybeans, washing the soybeans twice by using the deionized water, stirring the soybeans for 1min by using a soybean water ratio of 1/8 (the added water amount is × 9 percent of the mass of the dried soybeans-the mass of the soaked soybeans), grinding the soybeans into thick liquid, filtering the thick liquid by using 120-mesh gauze to obtain soybean milk, heating the soybean milk by using a boiling water bath, placing the heated soybean milk into an ice water bath to cool the soybean milk to room temperature after the final temperature of the soybean milk is 96 ℃, inoculating fermented soybean milk for 24 hours into the soybean milk according to a volume ratio of 10 percent, fermenting the soybean milk for 6 hours at 37 ℃, measuring the change of the pH value by using a pH meter, and measuring.
The results are shown in table 5, after 6h of fermentation, the pH value of lactobacillus plantarum CGMCC No.16112 can be reduced from 6.50 to 4.64, and compared with a natural fermentation group C1 and a lactobacillus paracasei fermentation group C2, the acid production capacity of the lactobacillus plantarum CGMCC No.16112 is similar to that of the natural fermentation group and is stronger than that of the lactobacillus paracasei, which indicates that the lactobacillus plantarum CGMCC No.16112 can ferment soybean milk to produce acid quickly and curd the soybean milk.
TABLE 5 pH in soymilk at different time points
Example seven: lactic acid production capacity of lactobacillus plantarum in soybean milk within 6h
Taking 1000m L fresh sterile soybean milk as raw material, adding 10m L yellow serofluid fermented for 24h, keeping the temperature at 37 ℃ for 6h, weighing 2m L samples, centrifuging at 10000rpm for 10min, filtering with 0.22 μm filter membrane, measuring the content of lactic acid, and performing three-time measurement.
As shown in Table 6, the lactic acid yield of 6.37 g/L can be obtained by fermenting the soybean milk in 6h with Lactobacillus plantarum CGMCC No.16112, which is 1.71-1.77 times that of the naturally fermented group C1 and the Lactobacillus paracasei fermented group C2, and the curd rate of the soybean milk is remarkably increased.
TABLE 6 lactic acid production in soymilk at different time points
Example eight: application of lactobacillus plantarum in sour soybean milk
Taking 1000m L fresh sterile soybean milk as a raw material, adding yellow serofluid of lactobacillus plantarum CGMCC No.16112 fermented for 24h, preserving heat for 6h at 37 ℃, weighing a sample, measuring the water binding capacity and strength value of the gel, and carrying out three times of measurement in parallel.
The results are shown in table 7, compared with the addition of 10% of fermented yellow pulp, the addition of 7% of fermented yellow pulp can make the bean curd form gel with better water retention rate and common strength, and the addition of 7% of lactobacillus plantarum CGMCC No.16112 is suitable for developing the sour soybean milk product with tender mouthfeel.
TABLE 7 Water holding Properties and Strength of Bean curd gels
Example nine: application of lactobacillus plantarum in bean curd production
After the activated lactobacillus plantarum CGMCC No.16112 is washed, the concentration of bacterial cells is adjusted to 109More than CFU/m L, freeze-drying to obtain direct vat set starter, inoculating 1g of bacteria powder into 1000m L soybean milk, fermenting at 37 deg.C for 4h, sampling to determine water holding rate, and measuring gel property with TA-XT Plus texture analyzer for three times.
As shown in Table 8, the water retention of the tofu fermented by Lactobacillus plantarum CGMCC No.16112 is better than that of the tofu fermented by natural lactobacillus paracasei, and the hardness, elasticity, cohesion and chewiness of the tofu gel are similar to those of the tofu fermented by natural lactobacillus paracasei and better than those of the tofu fermented by natural lactobacillus paracasei, which indicates that the tofu can be rapidly gelled and the texture of the tofu can be effectively improved by adding Lactobacillus plantarum CGMCC No. 16112.
TABLE 8 Water holding Properties and gel Properties of fermented sour Soybean milk with different strains
Example ten: application of lactobacillus plantarum in preparation of pickled vegetables
The method comprises the steps of placing cleaned radish raw materials into 10% sterile salt solution for pre-pickling for 24 hours, transferring radishes into low-concentration sterile salt solution for finished product pickling, collecting activated lactobacillus plantarum CGMCC No.16112, washing, inoculating 20m L viable bacteria liquid into 980m L finished product pickling liquid, pickling for 48 hours at 28 ℃, sampling, measuring pH and acidity of fermentation liquid, and measuring in parallel for three times.
The results are shown in table 9, lactobacillus plantarum CGMCC No.16112 can grow in salt water with different concentrations and can ferment the pickle liquor to produce acid quickly, when the concentration of the pickle liquor is 4%, the pickle is matured fastest, the crispness of the pickle can be effectively improved, and the quality of the pickle is better.
TABLE 9 pH and acidity values of finished pickling solutions of different concentrations
Example eleven: application of lactobacillus plantarum in yoghourt making
Using 980m L fresh skim milk as raw material, adding 4% white sugar, pasteurizing at 105 deg.C for 10min, inoculating 20m L bacteria with cell concentration of 106The activated bacterium liquid of Lactobacillus plantarum CGMCC No.16112 of CFU/m L is kept at 42 ℃ for 6h, and the curd condition (the curd condition is that the content does not flow when the vessel is inclined) and the acid production condition are observed.
As shown in Table 10, Lactobacillus plantarum CGMCC No.16112 can ferment in 6h to complete skim milk, stably and rapidly produce acid, and curd the skim milk to form typical yogurt texture.
TABLE 10 skim milk curdling at different time points
Note: "-" indicates that no curd was made
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
SEQUENCE LISTING
<110> university of south of the Yangtze river
<120> lactobacillus plantarum with high lactic acid yield and application thereof
<160>3
<170>PatentIn version 3.3
<210>1
<211>1435
<212>DNA
<213>16S rDNA
<400>1
tgcagtcgac gaactctggt attgattggt gcttgcatca tgatttacat ttgagtgagt 60
ggcgaactgg tgagtaacac gtgggaaacc tgcccagaag cgggggataa cacctggaaa 120
cagatgctaa taccgcataa caacttggac cgcatggtcc gagcttgaaa gatggcttcg 180
gctatcactt ttggatggtc ccgcggcgta ttagctagat ggtggggtaa cggctcacca 240
tggcaatgat acgtagccga cctgagaggg taatcggcca cattgggact gagacacggc 300
ccaaactcct acgggaggca gcagtaggga atcttccaca atggacgaaa gtctgatgga 360
gcaacgccgc gtgagtgaag aagggtttcg gctcgtaaaa ctctgttgtt aaagaagaac 420
atatctgaga gtaactgttc aggtattgac ggtatttaac cagaaagcca cggctaacta 480
cgtgccagca gccgcggtaa tacgtaggtg gcaagcgttg tccggattta ttgggcgtaa 540
agcgagcgca ggcggttttt taagtctgat gtgaaagcct tcggctcaac cgaagaagtg 600
catcggaaac tgggagactt gagtgcagaa gaggacagtg gaactccatg tgtagcggtg 660
aaatgcgtag atatatggaa gaacaccagt ggcgaaggcg gctgtctggt ctgtaactga 720
cgctgaggct cgaaagtatg ggtagcaaac aggattagat accctggtag tccataccgt 780
aaacgatgaa tgctaagtgt tggagggttt ccgcccttca gtgctgcagc taacgcatta 840
agcattccgc ctggggagta cggccgcaag gctgaaactc aaaggaattg acgggggccc 900
gcacaagcgg tggagcatgt ggtttaattc gaagctacgc gaagaacctt accaggtctt 960
gacatactat gcaaatctaa gagattagac gttcccttcg gggacatgga tacaggtggt 1020
gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac 1080
ccttattatc agttgccagc attaagttgg gcactctggt gagactgccg gtgacaaacc 1140
ggaggaaggt ggggatgacg tcaaatcatc atgcccctta tgacctgggc tacacacgtg 1200
ctacaatgga tggtacaacg agttgcgaac tcgcgagagt aagctaatct cttaaagcca 1260
ttctcagttc ggattgtagg ctgcaactcgcctacatgaa gtcggaatcg ctagtaatcg 1320
cggatcagca tgccgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca 1380
tgagagtttg taacacccaa agtcggtggg gtaacctttt aggaaccagc tgcta 1435
<210>2
<211>20
<212>DNA
<213> artificially synthesized sequence
<400>2
<210>3
<211>19
<212>DNA
<213> artificially synthesized sequence
<400>3
ggttaccttg ttacgactt 19
Claims (10)
1. The lactobacillus plantarum with high lactic acid yield is characterized in that the lactobacillus plantarum (L actinobacillus plantarum) is preserved in 16.07.2018 in China general microbiological culture Collection center (CGMCC), the preservation number is CGMCC No.16112, the preservation address is No. 3 of Xilu No.1 on North Chen of the sunward area in Beijing, and the institute of microbiology of China academy of sciences.
2. A microbial agent comprising the Lactobacillus plantarum strain defined in claim 1.
3. Use of the lactobacillus plantarum of claim 1 in the preparation of lactic acid.
4. Use of lactobacillus plantarum as claimed in claim 1 in the preparation of a soy product.
5. The use according to claim 4, wherein the Lactobacillus plantarum used in the preparation of a soy product comprises Lactobacillus plantarum used in the preparation of fermented soy milk and tofu.
6. Use of lactobacillus plantarum as defined in claim 1 in the preparation of kimchi.
7. Use of the lactobacillus plantarum of claim 1 in the manufacture of yoghurt.
8. Use of the microbial agent of claim 2 in the preparation of lactic acid.
9. Use of the microbial agent of claim 2 for the preparation of a bean product.
10. The use of the microbial inoculum of claim 2 in the preparation of pickles and yogurt.
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