CN109234348B - Anti-depression blood fat reducing monascus powder and preparation method thereof - Google Patents

Anti-depression blood fat reducing monascus powder and preparation method thereof Download PDF

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CN109234348B
CN109234348B CN201810682075.0A CN201810682075A CN109234348B CN 109234348 B CN109234348 B CN 109234348B CN 201810682075 A CN201810682075 A CN 201810682075A CN 109234348 B CN109234348 B CN 109234348B
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马伟超
李一婧
舒国伟
王廷璞
包文生
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Tianshui Normal University
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Abstract

The invention provides an anti-depression blood fat-reducing red rice powder and a preparation method thereof. Wherein the content of the hypolipidemic active ingredient lovastatin and the content of the antidepressant active ingredient GABA in the anti-depression hypolipidemic red yeast powder reach the highest values, which are respectively 0.22% (w/w) and 2.99% (w/w). The invention also provides a method for producing the anti-depression and blood fat-reducing monascus powder by fermenting and whole-cell catalytic mixed culture of monascus and lactic acid bacteria. The method can realize the synchronous promotion of lovastatin and GABA, and can be used for producing the anti-depression blood fat-reducing red yeast powder.

Description

Anti-depression blood fat reducing monascus powder and preparation method thereof
Technical Field
The invention relates to the field of microbial pharmacy, in particular to anti-depression blood fat-reducing monascus powder and a preparation method thereof.
Background
Red yeast rice, red koji or Fumi, is brown red to dark red, and is prepared from long-shaped rice, non-glutinous rice and glutinous rice through fermenting with red koji fungus(see Chensuangui, Zhang Dan, Liu Min, bear swallow, Zhang forest, Zhao Zi. high performance liquid chromatography for measuring lactone type and acid type lovastatin content in highland barley Monascus. Chinese brewing, 2016, (10):162-5 and Huangyan, Liyao Guanlong, Liu Si Xin. Monascus purpureus and safety research progress thereof. food research and development, 2006, (08): 217-21.). The main functional components in red yeast rice are Monascus pigment (see Shang Xiao, Zhang Fang, Zhang Ming, Li, Chen Sha, progress of research on biological activity of Monascus pigment, proceedings of Henan university of Industrial science (Nature edition), 2017 (02):129-35.), lovastatin (i.e., Monacolin K or mevinolin) (see Zhang J, Wang Y L, Lu L P, Zhang B, Xu G R. enhanced process of Monacolin K by addition of precursors and surfactants in synergistic reaction of Monascus purpureus 9901.Biotechnology and applied biochemistry, GABA, 61(2):202-7.), enzyme active substances (see chemistry-J, Wu-D, Chem I-S, 2011-7. and Tai Yu Ying-3. and Ying-6. amino acids of Chemicals-D. Shang-7. and Tai Yu-7. C. amino acids of Legend, 4. C. C.3. C.D. C.D.4. Ying-D.D.E.E.D.D.D.E.E.4. C.C.C. Yu.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.C.E.E.C.E.E.C.E.E.D.D.E.E.E.E.E.E.E.E.4. C.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.C.C.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E.E., shen X-H, Duan Z-W, Guo S-R.Advances on the Pharmacological Effects of Red Yeast Rice Rice.Chinese Journal of Natural Medicines,2011,9(3):161-6.), wherein lovastatin is an inhibitor of 3-Hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase which is the rate-limiting enzyme in the human cholesterol synthesis pathway, has the effect of reducing or blocking the synthesis of cholesterol in vivo, and thus has the efficacy of lowering blood pressure and lowering blood lipid (see Nakamura T, Komagata D, Murakawa S, Sakai K, Endo A. isolation and Biosynthesis of 3 α -Hydroxy-3, 5-Dihydromonatin L. the Journal of Antibiotics,1990,43(12):1597 and 1597, 600 and literature mirror, and routine, Journal of American and literature, and methods for determining physiological activities of lovastatin, 2001,112-9 and endo.A. Monacolin K, A New Hypocholesterol Agent Produced By A Monascus specifices the Journal of Antibiotics 1979,32(8): 852-4.). Another active ingredient, gamma-aminobutyric acid, is the major inhibitory neurotransmitter in the mammalian central nervous system (see Li H, Qiu T, Huang G, Cao Y. production of gamma-aminobutyric acid by Lactobacillus brevis NCL912using fed-batch transfer. Microbiologicalcell factors, 2010,9:85, Li K, Xu E, the role and the mechanism of gamma-aminobutyric acid reducing central nervous system devitalizing neural Bulletin,2008,24(3):195-200.), have the effects of reducing blood pressure, calming, inducing diuresis, and can prevent diabetes (see Li H, Qiu T, Gao D, Cao Y. medium optimization for development of gamma-aminobutyric acid by Lactobacillus brevis NC912. amino acids,2010,38(5):1439-45.)[12]. It has also been shown that GABA plays an important role in treating patients with depression (see Mann J, Oquesdo M A, Watson K T, Boldrin M, Malone K M, Ellis S P, Sullivan G, Cooper T B, Xie S, Currer D. Anxiety in major expression and cellulose flow from gamma-aminobutyl acid. expression and expression, 2014,31(10):814-21. and Pehrson A L, Sanchez C. organic gamma-amino acid circulation in major expression, a critical expression of lateral expression of GABA, a critical expression of the expression and expression of cellulose, intake of cellulose, moisture of expression of cellulose, intake of cellulose, intake of cellulose, cellulose. Therefore, the red yeast powder rich in lovastatin and GABA can have the dual effects of resisting depression and reducing blood fat.
Through recent studies, the content of lovastatin and GABA in red rice powder has been greatly increased, and it can be up to 0.59% (w/w) (see Jirased S, Nopharatana M, Kitsubu P, Vichisloonchul T, Tongta A. statistical optimization for Monacolin K and yellow pigment production and circulation reduction by Monascus purpureus in solid-state transfer. Journal of biological and biological technology,2013,23(3):364-74.) and 0.5% (w/w) (see Wang J, Lee C L, Pan T. M. improvement of Monacolin K, gamma-amino acid production of biological technology,2003, Journal of biological and biological technology, 76).
In addition, a 'Chinese food additive' published in 12.06.2011 published a paper of 'research on a process for producing gamma-aminobutyric acid by mixed fermentation of Monascus and Lactic acid bacteria', wherein the GABA produced by mixed fermentation of Monascus (Monascus) and Lactic acid bacteria (Lactic acid bacteria) has very obvious superiority and good development prospect. After the monascus SM048 and the plant lactobacillus Lac.1 are inoculated at the same time, the growth speed of the monascus is inhibited to a certain degree, and the monascus is suitable for mixed fermentation by adopting a segmented culture mode. Under the conditions of pH 4-4.5 and 30 ℃, the yield of GABA by stage fermentation is 0.52g/L, which is respectively improved by 147.62% and 62.5% compared with SM048 and Lac.1 which are fermented independently.
However, a fermentation method for simultaneously increasing the contents of two functional components has not been reported, and the taste of the red yeast powder produced by a single bacterial strain in the market is not good at present.
Disclosure of Invention
In order to effectively and simultaneously improve the contents of two functional components and the palatability of the red rice powder, the invention aims to provide the anti-depression blood fat-reducing red rice powder.
The second purpose of the invention is to provide a method for producing anti-depression and blood fat-reducing monascus powder by fermentation-whole cell catalysis mixed culture by utilizing monascus and lactic acid bacteria, the method produces functional monascus powder with synchronously improved lovastatin and GABA contents by carrying out mixed culture on a lovastatin high-yield monascus strain obtained by early screening and a GABA-producing lactic acid strain, and the lovastatin content in the monascus powder reaches 0.22% (w/w) and the GABA content reaches 2.99% (w/w). In addition, the addition of the lactic acid bacteria not only improves the GABA content in the red rice powder, but also provides various other beneficial components for the red rice powder and improves the mouthfeel of the red rice powder.
The anti-depression and blood fat reducing red rice powder as the first aspect of the invention contains the blood fat reducing active ingredient lovastatin and the anti-depression active ingredient GABA with the contents of 0.22% (w/w) and 2.99% (w/w), respectively.
The method for producing the anti-depression and blood fat reducing Monascus powder according to claim 1 by fermentation-whole cell catalytic mixed culture of Monascus and lactic acid bacteria according to the second aspect of the present invention is characterized in that Monascus (Monascus anka) Mon 20-2 after slant activation is inoculated to a seed culture medium for culture, and the Monascus fermentation broth is obtained by inoculating the Monascus after culture; adding activated lactobacillus into Monascus fermentation broth for whole-cell catalysis to obtain the anti-depression and blood lipid-lowering Monascus powder;
the Monascus purpureus (Monascus anka) Mon 20-2 is obtained by carrying parent Monascus purpureus 30192 (purchased from the China agricultural microbial culture collection management center, strain number ACCC 30192) on China eight spaceships, carrying out space mutagenesis, and screening, and is preserved in the China typical culture collection (China, Wuhan) at 6-4 days 2015 with the preservation number CCTCC M2015356.
In a preferred embodiment of the invention, the activated lactic acid bacteria are added into monascus fermentation liquor for conversion to obtain the anti-depression and blood fat-reducing monascus powder, and the dry weight ratio of monascus to lactic acid bacteria is 3-1: 1-3.
In a preferred embodiment of the invention, the lactic acid bacteria are Lactobacillus acidophilus (Lactobacillus acidophilus) L5, Lactobacillus bulgaricus (Lactobacillus bulgaricus) L41, Lactobacillus bulgaricus (Lactobacillus bulgaricus) L48, Lactobacillus rhamnosus (Lactobacillus rhamnosus) LD, Lactobacillus rhamnosus (Lactobacillus rhamnosus)7469, Lactobacillus rhamnosus (Lactobacillus rhamnosus) L22, Lactobacillus rhamnosus (Lactobacillus rhamnosus) L54, Lactobacillus rhamnosus (Lactobacillus rhamnosus) L5, Lactobacillus reuteri (Lactobacillus reuteri) L33, Lactobacillus reuteri (Lactobacillus reuteri) L21, Lactobacillus plantarum (Lactobacillus plantarum) L4834, Lactobacillus plantarum (Lactobacillus plantarum) L3874, Lactobacillus plantarum (Lactobacillus bifidus) L3625, Lactobacillus plantarum (Lactobacillus plantarum) L3625, Lactobacillus plantarum (Lactobacillus plantarum) L3625.
In a preferred embodiment of the invention, the conversion temperature of the anti-depression blood fat-reducing monascus powder obtained by adding the activated lactic acid bacteria into monascus fermentation liquor for conversion is 25-41 ℃.
In a preferred embodiment of the invention, the activated lactic acid bacteria are added into monascus fermentation liquor for conversion to obtain the ion concentration of the conversion system of the anti-depression blood fat-reducing monascus powder, wherein the ion concentration of the conversion system is 0.01 g/L-0.09 g/L.
In a preferred embodiment of the invention, the ion concentration is a zinc ion concentration.
In a preferred embodiment of the invention, the activated lactic acid bacteria are added into monascus fermentation liquor for conversion to obtain the antidepressant and blood fat-reducing monascus powder, and the pH value of the antidepressant and blood fat-reducing monascus powder is 3.5-7.5.
According to the invention, the lactobacillus strain and the monascus Mon 20-2 under the specific culture condition are introduced to combine to prepare the functional monascus powder with the lovastatin and GABA contents synchronously improved, the lovastatin and GABA contents are synchronously improved, the obtained monascus powder contains the aromatic odor of lactic acid bacteria, the bitter taste of monascus powder produced by single monascus fermentation is removed, and the monascus powder is more suitable for taking.
Drawings
FIG. 1a is a graph showing comparison of the conversion rate of L-glutamic acid in each strain in the preliminary screening test.
FIG. 1b is a graph showing a comparison of GABA production by each strain in the re-screening test.
FIG. 2 is a graph showing the comparison of glutamic acid conversion rate of Lactobacillus 8014 at different conversion temperatures.
FIG. 3a is a graph showing the comparison of the conversion rate of L-glutamic acid by lactic acid bacterium 8014 with different ions added.
FIG. 3b is a graph showing the comparison of the conversion rate of L-glutamic acid by Lactobacillus 8014 at various zinc ion concentrations.
FIG. 4 is a graph showing the comparison of glutamic acid conversion rate and GABA production of lactobacillus 8014 in different pH conversion systems.
FIG. 5 is a graph showing a comparison of Monacolin K and GABA production at different Monascus and Lactobacillus ratios.
Detailed Description
Materials and methods
1. Strain:
1.1 Monascus: monascus anka Mon 20-2 is obtained by screening parent strain Monascus 30192 (purchased from China agricultural microbial culture collection management center, strain number ACCC 30192) after being carried by China eight spaceship and subjected to space mutagenesis, and has been preserved in the China type culture collection (China, Wuhan) at 6-4 days in 2015 with the preservation number of CCTCC M2015356.
1.2 lactic acid bacterial strain: lactobacillus acidophilus (Lactobacillus acidophilus) L5; lactobacillus bulgaricus (Lactobacillus bulgaricus) L41, L48; lactobacillus rhamnosus LD, 7469, L22, L20, L57; lactobacillus reuteri (Lactobacillus reuteri) L33, L21; lactobacillus plantarum (Lactobacillus plantarum)8014, L69, L24; lactobacillus casei (Lactobacillus casei) L54, LC, L49; bifidobacterium longum (Bifidobacterium longum) B28. All the samples are screened and stored in the laboratory. Wherein Lactobacillus acidophilus L5 is isolated from strain CICC 6074 of China center for culture Collection of Industrial microorganisms; lactobacillus rhamnosus LD, L20, L22, L57 are respectively from China center strains CICC 20255, 20257, 20258, 20259; lactobacillus reuteri L21 and L33 are respectively derived from strains CICC 6118 and 6120 of China industrial microorganism culture collection management center; lactobacillus plantarum L24 and L69 are respectively derived from China center for Industrial culture Collection of microorganisms and strains CICC 20265 and 20267; lactobacillus casei (Lactobacillus casei) LC, L49, L54 are respectively derived from China center for culture Collection of Industrial microorganisms strains CICC 20280, 20282, 20284; bifidobacterium longum (Bifidobacterium longum) B28 is derived from strain CICC 6194 of China center for culture Collection of Industrial microorganisms; lactobacillus bulgaricus (Lactobacillus bulgaricus) L41, L48 was isolated from American type culture Collection strains ATCC 8001, 7517; lactobacillus rhamnosus 7469 was purchased from American type culture Collection (Strain No. ATCC 7469); lactobacillus plantarum (Lactobacillus plantarum)8014 was obtained from the American type culture Collection (strain No. ATCC 8014).
1.3 Medium
1.3.1 lactic acid bacteria culture Medium (MRS): 20.0g of glucose, 10.0g of beef extract, 10.0g of peptone, 5.0g of sodium acetate, 5.0g of yeast powder, 2.0g of diammonium citrate, 2.0g of disodium hydrogen phosphate, 801.0 g of tween, 0.1g of magnesium sulfate heptahydrate and 0.05g of manganese sulfate pentahydrate, dissolving in 1000ml of distilled water, sterilizing at 121 ℃ for 20 minutes, and separately sterilizing the glucose.
1.3.2 culture Medium of Monascus:
seed culture medium: dissolving 0.3% malt extract in distilled water, and sterilizing at 121 deg.C.
② fermentation medium: 7% of rice flour, 1.5% of ammonium chloride, 0.15% of dipotassium hydrogen phosphate and 0.05% of magnesium sulfate heptahydrate, and the pH value is natural.
1.4 screening of GABA-producing lactic acid bacteria
Respectively streak-inoculating lactobacillus strains stored at-80 deg.C in plate culture medium, culturing at 37 deg.C for 2d, transferring into shake tube, culturing at 37 deg.C in 250r/min shake table for 12h, and measuring OD600And (4) centrifuging to collect the bacteria. Inoculating activated lactic acid bacteria into 5mL of transformation system to final concentration OD600The transformation system included 6.25 g/L-glutamic acid, 0.1mM pyridoxal phosphate (PLP), CaCl2 50mM,MgSO450mM, acetate buffer (pH 4.8). After 12h of transformation, the residual concentration of L-glutamic acid in the transformation system was measured using an SBA-40E type biosensor analyzer to calculate the conversion rate of L-glutamic acid (R) of each strainGlu)。
The strains obtained by primary screening are inoculated into an MRS liquid culture medium, cultured for 12h at 37 ℃, inoculated into an MRS liquid culture medium containing 1g/L of L-glutamic acid in an inoculation amount of 1 percent, and cultured for 24h at 37 ℃. Centrifuging the fermentation liquid at 4 deg.C and 12000r/min for 15min, collecting supernatant, filtering with 0.45 μm filter membrane, and detecting the content of gamma-aminobutyric acid and Glu by High Performance Liquid Chromatography (HPLC).
1.5 Effect of temperature on the conversion of lactic acid bacteria to GABA
Inoculating Lactobacillus plantarum 8014 cultured overnight at an inoculum size of 1% into MRS liquid culture medium containing 1 g/L-glutamic acid, culturing in shaking table at 25 deg.C, 29 deg.C, 33 deg.C, 37 deg.C, 41 deg.C, and 250r/min, respectively, setting up 2 parallel groups in each of five groups, culturing for 24 hr, and measuring OD600Values and concentrations of Glu in the system.
1.6 Effect of ions on the conversion of lactic acid bacteria to GABA
(1) Preparing 10g/L calcium chloride, zinc sulfate, ferric chloride, ferrous chloride, cobalt chloride, magnesium sulfate, potassium dihydrogen phosphate and copper sulfate solution, adding into MRS culture medium at 0.5%, culturing at 33 deg.C for 24 hr as described in method 1.4, and detecting OD600Values and Glu concentrations.
(2) MRS culture media containing 0.01g/L, 0.03g/L, 0.05g/L, 0.07g/L and 0.09g/L zinc sulfate solutions are respectively prepared, and the influence of different zinc ion concentrations on the conversion and synthesis of GABA of lactobacillus plantarum 8014 is detected according to the conditions of the method 1.4.
1.7 Effect of pH on the conversion of lactic acid bacteria to GABA
Lactobacillus plantarum 8014 seed broth was inoculated at 5% inoculum size in a 3L fermentor containing 1L MRS medium (containing 0.05g/L zinc sulfate) and adjusted to pH 3.5, 4.5, 5.5, 6.5 and 7.5 with sodium hydroxide or hydrochloric acid, respectively. Bacteria solution OD600When 1.0 was reached, sodium glutamate was added to each tank to a final concentration of 1.15g/L (1 g/L relative to L-glutamic acid). After 24h of incubation, OD was determined600Value, concentration of Glu and GABA in the fermentation broth.
1.8 fermentation of Mixed strains to produce functional Red Rice
Inoculating Monascus ruber Mon 20-2 after slant activation to seed culture medium, culturing on shaking table at 28 deg.C and 200r/min for 48h, inoculating to 100mL medical saline bottle containing 40mL fermentation culture medium, and culturing on shaking table at 28 deg.C and 200r/min for 7 d. Lactobacillus 8014 cultured for 12h was centrifuged at 4000g for 5min at 4 ℃ and resuspended in phosphate buffer pH 7.5 as monascus: lactic acid bacteria (dry weight ratio) 3:1, 2:1, 1:2 and 1:3 are respectively added into monascus fermentation broth, L-glutamic acid solution is added until the final concentration is 1g/L, mixed fermentation culture is carried out in a shaking table at 33 ℃ and 200r/min, L-glutamic acid is supplemented to 1g/L every 20h, and each group is divided into 2 groups in parallel.
1.9 detection method
(1) Determination of dry weight of strain: taking the monascus liquid cultured for 7 days and the lactobacillus liquid cultured for 12 hours under the conditions of the method 1.8, centrifuging for 5min at 4 ℃ and 12000rpm, discarding the supernatant, drying to constant weight at 60 ℃, and weighing.
(2) L-glutamic acid and GABA were determined by Phenyl Isothiocyanate (PITC) pre-column derivatization, with specific reference to Roger (see Rogers K L, Philibert R A, Allen A J, Molitor J, Wilson E J, Dutton G R. HPLC analysis of reactive amino acids and neural residues. journal of neural methods,1987,22(2): 173-9.). The measurement of lovastatin and citrinin is in accordance with the national standard QB/T2847-2007 (refer to the Council of the people' S republic of China, the national development and reform Committee of QB/T2847-2007 functional Red Yeast Rice (powder) [ S ]. Beijing; China light industry Press, 2007.). HPLC detection Using a Thermo U-3000HPLC system, equipped with a DAD detector (DAD-3000), autosampler (WPS-3000SL) and fluorescence detector (FLD-3400 RS).
1.10 data analysis
The experimental data were analyzed using SPSS V20(IBM corporation), and the experimental condition data were tested using one-way analysis of variance (ANOVA) and Tukey Post Hoc. The chart was prepared using originPro 8.1.
2. Results and discussion
2.1 screening of GABA-producing lactic acid bacteria
Glutamic acid conversion ratio (R) to 17 lactic acid bacteriaGlu) The results of the tests are shown in FIG. 1 a. There was a significant difference in 12h L-glutamic acid conversion rate among the strains (p)<0.001 and alpha is 0.05), and the conversion rate of each strain is subjected to clustering Analysis (K-Means Cluster Analysis), so that the conversion rate of glutamic acid of the strains 8014, L48, LC, L57 and L5 is higher than that of other strains, and therefore 5 strains are selected and subjected to secondary screening by adding a precursor L-glutamic acid fermentation method. The rescreening results are shown in FIG. 1b, where there is a significant difference in GABA production (p) for each strain<0.001,. alpha. alpha.0.05), wherein the GABA yield of Lactobacillus plantarum 8014 is significantly higher than that of the other 4 strains, and the GABA content in the fermentation broth reaches 0.16g/L after culturing for 24h at 37 ℃ in an MRS liquid medium containing 1g/L of L-glutamic acid.
2.2 Effect of temperature on glutamic acid conversion
As shown in fig. 2, the conversion rate of glutamic acid was the highest in lactobacillus plantarum 8014 at 33 ℃, and the conversion rates of glutamic acid were significantly different among strains at different temperatures (p ═ 0.01, α ═ 0.05), indicating that the optimum L-glutamic acid conversion temperature of strain 8014 was 33 ℃. This temperature varies not only with the strain, such as the optimum transformation temperature of Lactobacillus plantarum Taj-Apis362 at 36 ℃ (see Tajabadi N, Baradaran A, Ebrahimoto A, Rahim R A, Bakar F A, Manap M Y, Mohammed A S, Saari N.Olexpression and optimization of phosphate dehydrogenase in Lactobacillus plantarum Taj-Apis362for high-gain-amino acid production. microbial biology, 2015,8(4):623-32.), while the optimum GABA transformation temperature of Lactobacillus brevis (L.brevis) TCCC13007 is at 30 ℃ (see Zhang Y, Song L, Gao Q, Yu S M, Li, o N.Thermoto-biological and 19. the optimum transformation temperature of Lactobacillus brevis 1616 ℃ (see Zhang Y, Song L, G Q, Yu S M, Li, G N.F.thermophil 3. gamma-biological and 9. biological analysis, 9. biological analysis and 19. 9. biological analysis; it also relates to the composition of the transformation system, such as the same strain Lactobacillus brevis (L.brevis) TCCC13007, which changes the optimal transformation temperature to 45 ℃ after changing the buffer composition and the substrate L-glutamic acid concentration in the transformation system (see Shi X, Chang C, Ma S, Cheng Y, Zhang J, Gao Q.Effective biological conversion of L-glutamate to gamma-aminobutyric acid by Lactobacillus brevis expressing cells, journal of industrial microbiology & biotechnology,2017,44(4-5): 697-).
2.3 Effect of ions on glutamic acid conversion
The effect of adding different ions in the transformation system on the L-glutamic acid transformation capability of Lactobacillus plantarum 8014 was examined, and the results are shown in FIG. 3 a. The addition of different ions significantly affected the L-glutamic acid-converting ability (p) of Lactobacillus plantarum 8014<0.001, α ═ 0.05), and has an effect on the growth of the strain, particularly when Mg is added alone2+Under the condition, the bacterial quantity is obviously reduced (p)<0.001, α ═ 0.05). To avoid the influence of the change of the amount of the bacteria on the conversion rate of the L-glutamic acid of the strain, the unit OD is determined by the test600L-glutamic acid conversion ratio (R) of cellsGlu/OD), the results show that Zn is contained in the solution2+The addition of (2) significantly enhances the transforming ability of L-glutamic acid. By the pair of Zn2+The concentration of (2) was optimized, and the result shows that the conversion rate of L-glutamic acid of Lactobacillus plantarum 8014 was the highest under the condition of adding 0.05g/L zinc sulfate (FIG. 3 b).
2.4 Effect of pH on Lactobacillus conversion to GABA
The L-glutamic acid conversion capability and GABA yield of Lactobacillus plantarum 8014 under different pH conditions were examined by using a mode of adjusting and controlling the pH of the conversion system in real time in a fermenter, and the results are shown in FIG. 4. After 24h of culture in MRS medium containing 1g/L L-glutamic acid, the conversion rate of L-glutamic acid is over 80% under each pH condition, and in a system with pH 7.5, the conversion rate of L-glutamic acid is obviously higher than that of other pH (p is 0.027, alpha is 0.05), the yield of GABA reaches 0.71g/L, and the conversion rate of L-glutamic acid reaches 100%.
2.5 fermentation of Mixed strains to produce functional Red Rice
In order to realize the synchronous promotion of lovastatin and GABA, the results of experiments on the yield of lovastatin and GABA by mixed fermentation of monascus and lactobacillus in different proportions by a two-stage method are shown in FIG. 5. Different monascus: under the condition of lactobacillus (dry weight ratio), the yields of lovastatin and GABA are remarkably different (p is less than 0.027, and alpha is 0.05), wherein when the dry weight ratio of monascus Mon 20-2 and lactobacillus plantarum 8014 is 2:1, the yields of the two functional components reach the highest values, namely 0.22% (w/w) and 2.99% (w/w), respectively. The mixed fermentation process obtained by the research can realize the synchronous promotion of lovastatin and GABA, the obtained monascus powder contains the aromatic odor of lactic acid bacteria, the bitter taste of monascus powder produced by single monascus fermentation is removed, and the monascus powder is more suitable for taking.
3. Conclusion
The invention develops a production process for producing anti-depression and blood fat-reducing monascus powder by fermenting and whole-cell catalytic mixed culture by utilizing monascus and lactic acid bacteria. In order to improve the content of an antidepressant functional component GABA, a GABA-producing strain lactobacillus plantarum 8014 is obtained by screening firstly in an experiment, and the optimum temperature, ions, pH and the like for converting L-glutamic acid are researched, and the result shows that the conversion rate of the L-glutamic acid can reach 100% in an MRS culture system with 0.05g/L zinc sulfate and pH of 7.5 added at 33 ℃. Then, the lactobacillus plantarum 8014 and the lactobacillus plantarum Mon 20-2 are subjected to fermentation-whole cell catalysis mixed culture according to different proportions, and the results show that when the lactobacillus plantarum Mon 20-2 and the lactobacillus plantarum 8014 are in a dry weight ratio of 2:1, the contents of the blood fat reducing active ingredient lovastatin and the antidepressant active ingredient GABA in the red koji powder obtained by fermentation reach the highest values, namely 0.22% (w/w) and 2.99% (w/w), respectively. The fermentation-whole cell catalysis mixed culture process obtained by the research can realize the synchronous promotion of lovastatin and GABA, the obtained monascus powder contains the aromatic odor of lactic acid bacteria, the bitter taste of monascus powder produced by single monascus fermentation is removed, and the monascus powder is more suitable for taking.

Claims (2)

1. A method for producing anti-depression and blood fat reducing Monascus powder by fermentation-whole cell catalytic mixed culture of Monascus and lactic acid bacteria is characterized in that Monascus (Monascus anka) Mon 20-2 after slant activation is inoculated to a seed culture medium for culture, and is inoculated to a fermentation culture medium for culture after culture to obtain Monascus fermentation liquor; adding activated lactobacillus into Monascus fermentation broth for conversion to obtain the anti-depression and blood fat-reducing Monascus powder;
the Monascus purpureus (Monascus anka) Mon 20-2 is obtained by carrying parent Monascus purpureus 30192 on China eight spaceship, carrying out space mutagenesis, and screening, and is preserved in the China center for type culture Collection (CCTCCM 2015356) at 6-4 months in 2015;
the activated lactic acid bacteria are added into monascus fermentation liquor for conversion to obtain the anti-depression and blood fat-reducing monascus powder, and the dry weight ratio of monascus to lactic acid bacteria is 3-1: 1-3;
the conversion temperature of the anti-depression blood fat-reducing monascus powder obtained by adding the activated lactic acid bacteria into monascus fermentation liquor for conversion is 25-41 ℃;
the activated lactobacillus is added into the monascus fermentation broth for conversion to obtain the ion concentration of 0.01-0.09 g/L in the conversion system of the antidepressant and blood fat reducing monascus powder;
the ion concentration is zinc ion concentration;
the activated lactic acid bacteria are added into monascus fermentation liquor for conversion to obtain the antidepressant and blood fat-reducing monascus powder, and the pH value of the antidepressant and blood fat-reducing monascus powder is 3.5-7.5;
the content of the hypolipidemic active ingredient lovastatin and the content of the antidepressant active ingredient GABA in the anti-depression hypolipidemic red yeast powder are respectively 0.22% (w/w) and 2.99% (w/w).
2. The method according to claim 1, wherein the lactic acid bacteria are Lactobacillus acidophilus (Lactobacillus acidophilus) L5, Lactobacillus bulgaricus (Lactobacillus bulgaricus) L41, Lactobacillus bulgaricus (Lactobacillus bulgaricus) L48, Lactobacillus rhamnosus (Lactobacillus rhamnosus) LD, Lactobacillus rhamnosus (Lactobacillus rhamnosus)7469, Lactobacillus rhamnosus (Lactobacillus rhamnosus) L22, Lactobacillus rhamnosus (Lactobacillus rhamnosus) L54, Lactobacillus rhamnosus (Lactobacillus rhamnosus) L57, Lactobacillus reuteri (Lactobacillus reuteri) L33, Lactobacillus reuteri (Lactobacillus casei) L638014, Lactobacillus rhamnosus (Lactobacillus rhamnosus) L57, Lactobacillus reuteri (Lactobacillus reuteri) L466, Lactobacillus plantarum (Lactobacillus plantarum) L48325, Lactobacillus plantarum (Lactobacillus bifidus) L3625, Lactobacillus plantarum (Lactobacillus plantarum) Lactobacillus).
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