CN105154492A - Liquid fermentation method for increasing yield of monascus polysaccharides - Google Patents
Liquid fermentation method for increasing yield of monascus polysaccharides Download PDFInfo
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Abstract
The invention relates to a liquid fermentation method for increasing the yield of monascus polysaccharides. The method includes the steps that a PCY liquid fermentation culture medium is inoculated with a monascus strain to perform fermentation culture so as to obtain a monascus seed solution, wherein the serial number of the monascus strain is CICC 40941 or CICC 40942 or CICC 40943 or CICC 40944, and the PCY liquid fermentation culture medium is prepared from 20 g of peptone, 20 g of maize meal, 10 g of yeast powder, 0.5 g of magnesium sulfate, 1.0 g of dipotassium phosphate, 0.1 g of ferrous sulfate and 1000 mL of distilled water; a PCY liquid culture medium is inoculated with the monascus seed solution according to the volume ratio (inoculum dose) of 1%-5% to perform fermentation culture so as to obtain a fermentation liquor and monascus mycelium mixture, and monascus polysaccharides are extracted from fermentation liquor which is separated out, wherein the PCY liquid culture medium contains rare earth elements (La3+ or Ce3+ or Pr3+ or Nd3+) with the concentration of 0.01-0.12 mmol/L. The method is simple in process, low in cost, significant in effect, environmentally friendly, nontoxic and capable of increasing the yield of monascus polysaccharides which can be used for developing immunity-improving drugs or anti-tumor drugs or anti-virus drugs.
Description
Technical field:
The present invention relates to a kind of liquid fermentation process utilizing rare earth element to improve monascus polysaccharide yield, belong to fermentation engineering field.
Background technology:
Monascus (
monascussp.) being China's traditional zymotic product---the production bacterial classification of red colouring agent for food, also used as a Chinese medicine, monascus is inoculated in the product of rice medium after fermentation and is red colouring agent for food, also used as a Chinese medicine, has the applicating history of more than one thousand years, be mainly used in food colorant, sanitas and drug matching in China.
Polysaccharide is the important secondary metabolite of one of monascus, research show monascus polysaccharide have antitumor, antiviral, improve the various biological effect such as immunizing power.At present, the research both at home and abroad about Fermentation Condition of Monascus spp product polysaccharide is less, and not yet realizes suitability for industrialized production.Existing research mainly improves monascus exopolysaccharides, as Zhao Zhenfeng etc. by fermentation condition optimization. and Fermentation Condition of Monascus spp produces the optimization of exocellular polysaccharide technique. Wuxi light industry college journal, 2002,2 (3): 289-295; Li Xiajiao etc. the optimization of Monascus polysaccharides liquid-state fermentation technology condition. biological processing, 2009,7 (2): 13-17; Wu Jianping etc. monascus liquid state fermentation polysaccharide process condition optimizing. Beijing Technology and Business University's journal (natural science edition) .2012,30 (6): 57-62.These researchs mainly concentrate in the technique aspects such as substratum composition, pH, temperature, dissolved oxygen and fermentation time, improve limited, can not meet the needs of modern industrialization fermentative production to monascus polysaccharide fermentation level.
Rare earth element is the general name of lanthanon and two coherent element scandiums and yttrium in the periodic table of chemical element, and China's rare earth resources enriches.At present, rare earth element is widely used in every field, is not only applied in each industrial sector, also increasing in the applied research of the field of biology such as agricultural, medicine, microorganism.The research of the microbial fermentation product of APPLICATION OF RARE-EARTH ELEMENTS raising in recent years has been reported, as Liu Yanmao etc. and rare earth element is on the impact of abomacetin fermentation. industrial microorganism, 2009,39 (1): 16-21; Li Zengli. Rare Earth Elements of Praseodymium, neodymium, erbium are on the impact of Production of Polysaccharide by Submerged Fermentation of Russula sp. Food science, 2007,28 (12): 312-316; And Wang Yi equality. the growth of rare earth element on rhodotorula and the impact of carotenogenesis. microbiology is circulated a notice of, and 1999,26 (2): 117-119.At present, there is not been reported in the applied research of rare earth element on monascus polysaccharide liquid fermenting.
Summary of the invention:
The object of the invention is to overcome the deficiency of above-mentioned prior art and provide that a kind of technique is simple, with low cost, the rare earth element that utilizes of Be very effective, environment-protecting asepsis improves the liquid fermentation process of monascus polysaccharide yield, present method can be used for the suitability for industrialized production of monascus polysaccharide.The present invention aims to provide the application of rare earth element in monascus polysaccharide liquid fermenting, provides the using method of rare earth element in monascus polysaccharide fermentation simultaneously, thus provides present method.
Object of the present invention can be reached by following measure: a kind of liquid fermentation process improving monascus polysaccharide yield, is characterized in that it comprises the steps:
(1) be inoculated in PCY liquid fermentation medium by monascus specie and carry out fermentation culture, culture condition is shaking speed 100 ~ 180r/min, temperature 25 ~ 30 DEG C, and incubation time 4 ~ 8 days, obtains monascus seed liquor;
Described monascus specie be monascus parpureus Went (
monascuspurpureus), bacterium numbering is CICC40941, CICC40942, CICC40943 or CICC40944;
Described PCY liquid fermentation medium consists of peptone 20g, Semen Maydis powder 20g, yeast powder 10g, magnesium sulfate 0.5g, dipotassium hydrogen phosphate 1.0g, ferrous sulfate 0.1g and distilled water 1000mL;
(2) it is in the PCY liquid nutrient medium of 0.01 ~ 0.12mmol/L rare earth element that monascus seed liquor step (1) obtained is linked into containing concentration by 1% ~ 5% volume ratio inoculum size, carry out fermentation culture, culture condition is 100 ~ 180r/min, temperature 25 ~ 30 DEG C, 4 ~ 8 days time, obtain fermented liquid and Monascus mycelium mixture, isolate fermented liquid for subsequent use;
Described rare earth element is lanthanum La
3+, cerium Ce
3+, praseodymium Pr
3+or neodymium Nd
3+;
(3) from the fermented liquid that step (2) obtains, monascus polysaccharide is extracted.
Monascus polysaccharide yield can reach 2.86g/L, adds lanthanum La
3+, cerium Ce
3+, praseodymium Pr
3+or neodymium Nd
3+after, monascus polysaccharide yield can reach respectively: 4.29g/L, 4.70g/L, 4.13g/L and 4.50g/L.
According to aforesaid method, the present invention does preferably following further:
Rare earth element described in step (2) is preferably cerium Ce
3+or neodymium Nd
3+, most preferred rare earth element is cerium Ce
3+.
The concentration of rare earth element in PCY liquid nutrient medium described in step (2) is preferably cerium Ce
3+0.04 ~ 0.08mmol/L, neodymium Nd
3+0.01 ~ 0.08mmol/L, lanthanum La
3+0.04 ~ 0.12mmol/L or praseodymium Pr
3+0.01 ~ 0.04mmol/L.
Described in step (2) is that the PCY liquid nutrient medium of 0.01 ~ 0.12mmol/L rare earth element is prepared by the following method containing concentration:
Take purity be 99% cerous nitrate, praseodymium nitrate, neodymium nitrate or lanthanum nitrate, be mixed with distilled water the solution that concentration is 1mol/L; Join in described PCY liquid fermentation medium, obtain and contain the PCY liquid nutrient medium that concentration is 0.01 ~ 0.12mmol/L rare earth element.
Fermented liquid described in step (2) and the separation of Monascus mycelium are through 2 layers of tulle silk (or 8 layers of gauze) filtering separation.
The extraction step of the monascus polysaccharide described in step (3) is as follows:
Fermented liquid is settled to 200ml, gets fermented liquid 20ml and add 80ml dehydrated alcohol, hold over night at 5 DEG C, the centrifugal 10min of 5000r/min, gained precipitation is weighed after 60 DEG C of dryings.
Gained monascus polysaccharide of the present invention can be used for the exploitation improving immunizing power, the medicine such as antitumor, antiviral.
The present invention compared with the prior art tool has the following advantages:
1, the present invention is by rare-earth element cerium Ce
3+, praseodymium Pr
3+, neodymium Nd
3+or lanthanum La
3+for the liquid fermenting of monascus polysaccharide, increase substantially the output of monascus polysaccharide, polysaccharide of fermentation broth output can reach 4.7g/L, has good industrial applications prospect.
2, rare-earth element cerium Ce of the present invention
3+, praseodymium Pr
3+, neodymium Nd
3+or lanthanum La
3+wide material sources, all have good facilitation effect, experimental technique is easy, reproducible.
3, monascus liquid fermentation process of the present invention is simple, environment-protecting asepsis, and material cost is cheap, and the whole fermenting process of what is more important is controlled, not by external environment condition restriction, is very applicable to suitability for industrialized production.
Accompanying drawing illustrates:
Fig. 1 is the influence curve figure of rare earth element to monascus polysaccharide yield.
Embodiment:
Below in conjunction with embodiment, the specific embodiment of the present invention is elaborated, raw materials used and substratum is as follows:
Fermentation monascus specie be monascus parpureus Went (
monascuspurpureus), bacterium numbering is CICC40941, CICC40942, CICC40943 or CICC40944, purchased from Chinese industrial Microbiological Culture Collection administrative center.Purity be 99% cerous nitrate, neodymium nitrate, lanthanum nitrate and praseodymium nitrate, purchased from Tianjin recovery fine chemistry industry institute.
PCY liquid fermentation medium consists of peptone 20g, Semen Maydis powder 20g, yeast powder 10g, magnesium sulfate 0.5g, dipotassium hydrogen phosphate 1.0g, ferrous sulfate 0.1g and distilled water 1000mL.
The preparation of the PCY liquid nutrient medium containing rare earth element:
Take purity be 99% cerous nitrate, praseodymium nitrate, neodymium nitrate or lanthanum nitrate, the solution that concentration is 1mol/L is mixed with distilled water, join in described PCY liquid fermentation medium, obtain respectively: (1) is 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L rare-earth element cerium Ce containing concentration
3+pCY liquid nutrient medium; (2) be 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L Rare Earth Elements of Praseodymium Pr containing concentration
3+pCY liquid nutrient medium; (3) be 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L rear earth element nd Nd containing concentration
3+pCY liquid nutrient medium; (4) be 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L rare-earth elements of lanthanum La containing concentration
3+pCY liquid nutrient medium for subsequent use.
Embodiment 1: a kind of liquid fermentation process improving monascus polysaccharide yield, step is:
1. carry out fermentation culture by monascus parpureus Went CICC40941 strain inoculation to PCY liquid fermentation medium, culture condition is shaking speed 160r/min, temperature 25 DEG C, and incubation time 8 days, obtains monascus seed liquor.
2. being linked into containing concentration by the monascus seed liquor of above-mentioned acquisition by 1% volume ratio inoculum size is 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L rare-earth element cerium Ce
3+pCY liquid nutrient medium in, carry out fermentation culture, culture condition is 160r/min, temperature 25 DEG C, 8 days time, obtains fermented liquid and Monascus mycelium mixture, filters to isolate fermented liquid through 2 layers of tulle silk, extracts monascus polysaccharide.
3. the extraction of monascus polysaccharide: fermented liquid is settled to 200ml, gets fermented liquid 20ml and adds 80ml dehydrated alcohol, hold over night at 5 DEG C, the centrifugal 10min of 5000r/min, and gained precipitation is weighed after 60 DEG C of dryings.Result as shown in Figure 1.
Embodiment 2: a kind of liquid fermentation process improving monascus polysaccharide yield, step is:
1. carry out fermentation culture by monascus parpureus Went CICC40942 strain inoculation to PCY liquid fermentation medium, culture condition is shaking speed 150r/min, temperature 30 DEG C, and incubation time 7 days, obtains monascus seed liquor.
2. being linked into containing concentration by the monascus seed liquor of above-mentioned acquisition by 2% volume ratio inoculum size is 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L Rare Earth Elements of Praseodymium Pr
3+pCY liquid nutrient medium in, carry out fermentation culture, culture condition is 150r/min, temperature 30 DEG C, and incubation time 7 days obtains fermented liquid and Monascus mycelium mixture, isolates fermented liquid through 8 layers of filtered through gauze, extracts monascus polysaccharide.
3. the extraction of monascus polysaccharide: fermented liquid is settled to 200ml, gets fermented liquid 20ml and adds 80ml dehydrated alcohol, hold over night at 5 DEG C, the centrifugal 10min of 5000r/min, and gained precipitation is weighed after 60 DEG C of dryings.Result as shown in Figure 1.
Embodiment 3: a kind of liquid fermentation process improving monascus polysaccharide yield, step is:
1. carry out fermentation culture by monascus parpureus Went CICC40943 strain inoculation to PCY liquid fermentation medium, culture condition is shaking speed 170r/min, temperature 28 DEG C, and incubation time 6 days, obtains monascus seed liquor.
2. being linked into containing concentration by the monascus seed liquor of above-mentioned acquisition by 5% volume ratio inoculum size is 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L rear earth element nd Nd
3+pCY liquid nutrient medium in, carry out fermentation culture, culture condition is 170r/min, temperature 28 DEG C, and incubation time 4 days obtains fermented liquid and Monascus mycelium mixture, filters to isolate fermented liquid through 2 layers of tulle silk, extracts monascus polysaccharide.
3. the extraction of monascus polysaccharide: fermented liquid is settled to 200ml, gets fermented liquid 20ml and adds 80ml dehydrated alcohol, hold over night at 5 DEG C, the centrifugal 10min of 5000r/min, and gained precipitation is weighed after 60 DEG C of dryings.Result as shown in Figure 1.
Embodiment 4: embodiment 1: a kind of liquid fermentation process improving monascus polysaccharide yield, step is:
1. carry out fermentation culture by monascus parpureus Went CICC40944 strain inoculation to PCY liquid fermentation medium, culture condition is shaking speed 180r/min, temperature 30 DEG C, and incubation time 4 days, obtains monascus seed liquor.
2. being linked into containing concentration by the monascus seed liquor of above-mentioned acquisition by 1% volume ratio inoculum size is 0.01mmol/L, 0.04mmol/L, 0.08mmol/L, 0.12mmol/L rare-earth elements of lanthanum La
3+pCY liquid nutrient medium in, carry out fermentation culture, culture condition is 180r/min, temperature 30 DEG C, and incubation time 6 days obtains fermented liquid and Monascus mycelium mixture, through or 8 layers of filtered through gauze isolate fermented liquid, extract monascus polysaccharide.
3. the extraction of monascus polysaccharide: fermented liquid is settled to 200ml, gets fermented liquid 20ml and adds 80ml dehydrated alcohol, hold over night at 5 DEG C, the centrifugal 10min of 5000r/min, and gained precipitation is weighed after 60 DEG C of dryings.Result as shown in Figure 1.
Above embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various distortion that the common engineering technical personnel in this area make technical scheme of the present invention and improvement, all should fall within protection domain that claims of the present invention determine.
Claims (7)
1. improve a liquid fermentation process for monascus polysaccharide yield, it is characterized in that it comprises the steps:
(1) be inoculated in PCY liquid fermentation medium by monascus specie and carry out fermentation culture, culture condition is shaking speed 100 ~ 180r/min, temperature 25 ~ 30 DEG C, and incubation time 4 ~ 8 days, obtains monascus seed liquor;
Described monascus specie be monascus parpureus Went (
monascuspurpureus), bacterium numbering is CICC40941, CICC40942, CICC40943 or CICC40944;
Described PCY liquid fermentation medium consists of peptone 20g, Semen Maydis powder 20g, yeast powder 10g, magnesium sulfate 0.5g, dipotassium hydrogen phosphate 1.0g, ferrous sulfate 0.1g and distilled water 1000mL;
(2) it is in the PCY liquid nutrient medium of 0.01 ~ 0.12mmol/L rare earth element that monascus seed liquor step (1) obtained is linked into containing concentration by 1% ~ 5% volume ratio inoculum size, carry out fermentation culture, culture condition is 100 ~ 180r/min, temperature 25 ~ 30 DEG C, 4 ~ 8 days time, obtain fermented liquid and Monascus mycelium mixture, isolate fermented liquid for subsequent use;
Described rare earth element is lanthanum La
3+, cerium Ce
3+, praseodymium Pr
3+or neodymium Nd
3+;
(3) from the fermented liquid that step (2) obtains, monascus polysaccharide is extracted.
2. a kind of liquid fermentation process improving monascus polysaccharide yield according to claim 1, is characterized in that described rare earth element is cerium Ce
3+or neodymium Nd
3+.
3. a kind of liquid fermentation process improving monascus polysaccharide yield according to claim 1 and 2, is characterized in that described rare earth element is cerium Ce
3+.
4. a kind of liquid fermentation process improving monascus polysaccharide yield according to claim 1, is characterized in that the described concentration of the rare earth element described in step (2) in PCY liquid nutrient medium is cerium Ce
3+0.04 ~ 0.08mmol/L, neodymium Nd
3+0.01 ~ 0.08mmol/L, lanthanum La
3+0.04 ~ 0.12mmol/L or praseodymium Pr
3+0.01 ~ 0.04mmol/L.
5. a kind of liquid fermentation process improving monascus polysaccharide yield according to claim 1, is characterized in that described in described step (2) being that the PCY liquid nutrient medium of 0.01 ~ 0.12mmol/L rare earth element is prepared by the following method containing concentration:
Take purity be 99% cerous nitrate, praseodymium nitrate, neodymium nitrate or lanthanum nitrate, be mixed with distilled water the solution that concentration is 1mol/L; Join in described PCY liquid fermentation medium, obtain and contain the PCY liquid nutrient medium that concentration is 0.01 ~ 0.12mmol/L rare earth element.
6. a kind of liquid fermentation process improving monascus polysaccharide yield according to claim 1, is characterized in that the separation of the described fermented liquid described in step (2) and Monascus mycelium is through 2 layers of tulle silk (or 8 layers of gauze) filtering separation.
7. a kind of liquid fermentation process improving monascus polysaccharide yield according to claim 1, is characterized in that the extraction step of the described monascus polysaccharide described in step (3) is as follows:
Fermented liquid is settled to 200ml, gets fermented liquid 20ml and add 80ml dehydrated alcohol, hold over night at 5 DEG C, the centrifugal 10min of 5000r/min, gained precipitation is weighed after 60 DEG C of dryings.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109609466A (en) * | 2019-01-24 | 2019-04-12 | 天津瑞生物科技股份有限公司 | The anti-aspergillus granulose hybridoma cell strain of mouse, monoclonal antibody and application |
| CN112094955A (en) * | 2020-09-15 | 2020-12-18 | 淄博职业学院 | Method for preparing oligosaccharide from pure polysaccharide |
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| CN102277303A (en) * | 2011-08-08 | 2011-12-14 | 浙江师范大学 | Monascus rubber Mr-99 strain and application thereof |
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2015
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| CN101880700A (en) * | 2010-07-20 | 2010-11-10 | 山东省农业科学院土壤肥料研究所 | Liquid fermentation method capable of improving yield of ganoderma iucidum polysaccharide |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109609466A (en) * | 2019-01-24 | 2019-04-12 | 天津瑞生物科技股份有限公司 | The anti-aspergillus granulose hybridoma cell strain of mouse, monoclonal antibody and application |
| CN109609466B (en) * | 2019-01-24 | 2022-03-01 | 天津一瑞生物科技股份有限公司 | Mouse aspergillus polysaccharide hybridoma cell strain, monoclonal antibody and application |
| CN112094955A (en) * | 2020-09-15 | 2020-12-18 | 淄博职业学院 | Method for preparing oligosaccharide from pure polysaccharide |
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Application publication date: 20151216 |