CN102757905B - Kitchen waste HBS-Hab(pichia pastoris) and application thereof - Google Patents
Kitchen waste HBS-Hab(pichia pastoris) and application thereof Download PDFInfo
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- CN102757905B CN102757905B CN2012101100864A CN201210110086A CN102757905B CN 102757905 B CN102757905 B CN 102757905B CN 2012101100864 A CN2012101100864 A CN 2012101100864A CN 201210110086 A CN201210110086 A CN 201210110086A CN 102757905 B CN102757905 B CN 102757905B
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Abstract
The invention relates to kitchen waste HBS-Hab(pichia pastoris), of which the preservation number is CGMCC No.5960. The HBS-Hab(pichia pastoris) has a high-temperature fermentation capability, can fast decompose lactic acid, amino acid, organism, protein and other matrixes at the temperature of 60 DEG C, has a higher breading capability than other yeasts, and can coexist with lactobacillus so as to be significant to promote the appreciation of the lactobacillus; furthermore, the HBS-Hab(pichia pastoris) is mainly in charge of the high-temperature fermentation of the kitchen waste to accelerate the decomposition rate and promote the appreciation of effective microorganisms. The HBS-Hab(pichia pastoris) belongs to non-toxic strains, and has an obvious aerobic treatment effect on the kitchen waste and a lower treatment cost, thereby having a great practical and population value.
Description
Technical field
The invention belongs to field of microbial biotechnology, particularly a Yeasts and application thereof.
Background technology
The changing food waste main component comprises rice and flours food residues, vegetables, vegetable and animals oils, meat bone etc., on chemical constitution, starch, Mierocrystalline cellulose, protein, lipid and inorganic salt is arranged.Principal feature is that organic content is abundant, moisture content is high, perishable, and its proterties and smell all can make a very bad impression to environmental health, and easily grow pathogenic micro-organism, the objectionable impurities such as mycotoxin.
Due to cooking culture and the custom of having a dinner party, produce the changing food waste of flood tide every day.The annual changing food waste that produces of Chinese city is not less than 6,000 ten thousand tons.Nutritious changing food waste is valuable renewable resources.But owing to not yet drawing attention, method of disposal is improper, and it has become affects the potentially dangerous of food safety and ecological safety source.Changing food waste has the dual nature of refuse and resource, can be described as typical " having misplaced local resource ".And unprocessed directly raising livestock and poultry can bring harm to HUMAN HEALTH by the accumulation of livestock and poultry vivotoxin, objectionable impurities again, thereby cause the cross infection between people and animals.Contain the toxic substances such as flavacin, benzene in " sewer oil " that also has present illegal retailer to sell, get back to people's dining table through underground approach, human consumption causes the generation of chronic disease even carcinogenic, and people healthy caused great harm.
On September 22nd, 2011, Standing Committee of Beijing Municipal People's Congress's review " Domestic Waste In Beijing management rules (draft) ".Be to cut off sewer oil and flow into the channel of dining table, draft requires that certain scale is qualified should build rubbish treatment facility on the spot with regard to the meal.The water content of changing food waste is high, and easily corrupt, stink is large, easily goes into secondary pollution, the inconvenience of also bringing for its transportation.Therefore, how to carry out the original position harmless treatment of changing food waste to continuous effective fast, become the current social problem demanding prompt solution.In recent years, in the changing food waste Aerobic, because changing food waste has easy corruption, becomes sour, cause the environment for the treatment of media very easily to change, make the microbic activity capacity of decomposition descend, the smell and the treatment effect that produce stench undesirable thereupon.
Existing to changing food waste processing microbial inoculum, mostly adopt the conducts such as genus bacillus, pseudomonas to clear up bacterial strain, because these bacterial classification resistance to acids are not strong, the ability that suppresses the harmful microorganism growth, cause the treatment time short, process filler and add the easy step-down of pH in process at changing food waste, cause the microorganism growth environmental change, capacity of decomposition descends, and treatment effect is undesirable.
Summary of the invention
In order to address the above problem, the object of the present invention is to provide a kind of changing food waste to subdue the type yeast.
An also purpose of the present invention is to provide a kind of changing food waste to subdue the application of type yeast in decomposing changing food waste.
In order to realize the object of the invention, the present invention also provides a kind of changing food waste to subdue type yeast HBS-Hab (Pichia pastoris), and its preserving number is CGMCC No.5960.
Described preserving number is that the yeast of CGMCC No.5960 has the thermophilic fermentation ability, can be at the temperature of 60 ℃ the matrix such as fast decoupled lactic acid, amino acid, organism, protein, compare other yeast fecundities stronger, this bacterium can coexist with milk-acid bacteria, to promoting milk-acid bacteria increment tool to have very important significance, this bacterium mainly is responsible for the thermophilic fermentation of changing food waste, accelerates rate of decomposition, promotes the increment of effective microbe.Described bacterial classification is spherical bacterium, and bacterium colony is oyster white on wort agar, tarnish, and there is thin breach at the edge.Cultivate in wort, the nutrient solution surface has white and coarse bacterium uncut jade wrinkle has bacterial sediment at the end.
A kind of changing food waste of the present invention is subdued type milk-acid bacteria HBS-RS (Pediococcus acidilactici), and its preserving number is CGMCC No.5959.
Described preserving number is that the milk-acid bacteria of CGMCC No.5959 can decomposition glucose and protein, it is transformed into lactic acid more than 50%, therefore suitablely breed under sour environment, and has a very strong capacity antacid, can survive in the pH value is 3 environment, compare other milk-acid bacterias and have more acid resistance.
Above-mentioned milk-acid bacteria is spherical shape, and two planes alternately divide formation tetrad shape at the right angle, and general cell is given birth in pairs, and single survivor is rare, does not become catenation.Gram-positive is not moved, and produces acid, amphimicrobian.Bacterium colony is little on the MRS substratum, is white in color.Grower along agar puncture line is thread.Catalase is negative, does not produce cytopigment.
On the other hand, the present invention also provides a kind of changing food waste to subdue the type complex microbial inoculum, it comprises the component of following weight percent: milk-acid bacteria (Pediococcus acidilactici) 0.5%-2%, yeast (Pichia pastoris) 0.5%-1.5%, rice bran 40%-50%, wheat bran 40-50%, beans cypress 1%-10%, protein powder 1%-10%; The effective content of described milk-acid bacteria is 1.5 * 10
6-3.0 * 10
6Cfu/g, the effective content of described yeast are 1.0 * 10
6-3.5 * 10
6Cfu/g.
Also on the one hand, the invention provides a kind of changing food waste and subdue the application of type yeast in decomposing changing food waste.
Beneficial effect of the present invention:
Yeast provided by the invention has the thermophilic fermentation ability, can be at the temperature of 60 ℃ the matrix such as fast decoupled lactic acid, amino acid, organism, protein, compare other yeast fecundities stronger, this bacterium can coexist with milk-acid bacteria, to promoting milk-acid bacteria increment tool to have very important significance, this bacterium mainly is responsible for the thermophilic fermentation of changing food waste, accelerates rate of decomposition, promotes the increment of effective microbe.Saccharomycodes of the present invention to changing food waste aerobic treatment successful, reduces processing cost, so very large practical and popularizing value is arranged in non-toxic strain.
Description of drawings
Fig. 1 is that composite fungus agent of the present invention is processed 60 days garbage weight variation diagrams;
Fig. 2 is that yeast of the present invention is processed 30 days garbage weight variation diagrams.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, modification or replacement to the inventive method, step or condition are done all belong to protection scope of the present invention.
If do not specialize, the conventional means that in embodiment, technique means used is well known to those skilled in the art.
Embodiment 1
Described milk-acid bacteria and yeast all separate acquisition from swill, the invention discloses a kind of milk-acid bacteria (Pediococcus acidilactici), and its preservation name is called HBS-RS, preserving number: CGMCC No.5959.The invention also discloses a kind of pichia pastoris (Pichia pastoris), its preservation name is called HBS-Hab, preserving number: CGMCC No.5960.Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica; Preservation date: on April 9th, 2012.
Embodiment 2 changing food wastes are subdued the type complex microbial inoculum
1) slant culture: will be inoculated in respectively on solid medium under the yeast in embodiment 1 and milk-acid bacteria original strain aseptic condition, cultivated 2 days under 37 ℃ of conditions;
2) be inoculated in liquid nutrient medium under the bacterial classification aseptic condition of first order seed cultivation: with step 1) cultivating, under 42 ℃ of conditions, the 150r/min shaking table was cultivated 1 day, yeast and milk-acid bacteria suspension liquid light density OD when finishing to cultivate
600Value all reaches 4.0;
3) secondary seed is cultivated: be 10% inoculum size by the volume ratio of liquid nutrient medium, first order seed is inoculated into respectively in the fermentor tank of 1000L, in fermentor tank, the cumulative volume of nutrient solution is 500L, under 42 ℃ of conditions of yeast and milk-acid bacteria, stirring velocity is 150r/min, air flow is 1: 1, cultivates 1 day, makes secondary seed;
Wherein, step 1), 2), yeast and milk-acid bacteria substratum used are proteolytic enzyme peptone (protease peptone) 1g, beef extract (beef extract) 1g, glucose (glucose) 2g, sodium acetate (sodium acetate) 3g, and Trisodium Citrate (ammoni acitrate) 2g.
The formula of substratum step 3) used is by mass percentage: Semen Maydis powder 2.5%, soyflour 1%, peptone 1%, yeast powder 2.5%, K
2HPO
40.5%, MgSO
40.005%, tween 80 1%, sodium-acetate 0.5%, surplus are water, pH6.4.
The fermentation culture process comprises: in initial 12 hours, the interval ventilation remains on the aerobic conditions fermentation, air flow 1: 1, and regulation and control fermentation dissolved oxygen 10%, mixing speed 180r/min, stirred 42 ℃ of temperature 2 minutes at 2 hours mixing chamber intervals; Slightly soluble oxygen and anaerobism cultivation stage: after 12 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation, stir at the interval, 4 hours mixing chamber intervals, stirred 42 ℃ of temperature 4 minutes.
Add solid fermentation substratum 7500kg in 500 liters of numerous cultivation bacterium liquid of expansion, described solid fermentation substratum is milk-acid bacteria Pediococcus acidilactici1.5%, yeast Pichia pastoris 1%, rice bran (46%), wheat bran (45.5%), beans cypress (5%), protein powder (1%) stir, use the film capping, every stirring in 24 hours, cultivated 72 hours.
Embodiment 3
Expand numerous bacterial classification according to liquid fermentation process in example 2.
Wherein, step 1), 2), yeast and milk-acid bacteria substratum used are proteolytic enzyme peptone (protease peptone) 1.5g, beef extract (beef extract) 0.5g, glucose (glucose) 1g, sodium acetate (sodium acetate) 3g, and Trisodium Citrate (ammoni acitrate) 2g.
The formula of substratum step 3) used is by mass percentage: Semen Maydis powder 1%, soyflour 1%, peptone 1%, yeast powder 1.5%, K
2HPO
40.5%, MgSO
40.005%, tween 80 1%, sodium-acetate 0.5%, surplus are water, pH6.4.
The fermentation culture process comprises: in initial 12 hours, the interval ventilation remains on the aerobic conditions fermentation, air flow 1: 1, and regulation and control fermentation dissolved oxygen 10%, mixing speed 180r/min, stirred 41 ℃ of temperature 2 minutes at 2 hours mixing chamber intervals; Slightly soluble oxygen and anaerobism cultivation stage: after 12 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation, stir at the interval, 4 hours mixing chamber intervals, stirred 41 ℃ of temperature 4 minutes.
Expand numerous cultivation bacterium liquid with 500 liters and join (7500kg-8000kg) in the solid fermentation substratum.Described solid fermentation substratum is milk-acid bacteria Pediococcus acidilactici1.5%, yeast Pichia pastoris1%, rice bran (40%), wheat bran (40%), beans cypress (7.5%), protein powder (10%), stir, use the film capping, every stirring in 24 hours, cultivated 48 hours.
Embodiment 4
Expand numerous bacterial classification according to liquid fermentation process in example 2.
Wherein, step 1), 2), yeast and milk-acid bacteria substratum used are proteolytic enzyme peptone (protease peptone) 1g, beef extract (beef extract) 1g, glucose (glucose) 2g, sodium acetate (sodium acetate) 3g, and Trisodium Citrate (ammoni acitrate) 2g.
The formula of substratum step 3) used is by mass percentage: Semen Maydis powder 2.5%, soyflour 2.5%, peptone 2%, yeast powder 2.5%, K
2HPO
41.5%, MgSO
40.005%, tween 80 1%, sodium-acetate 0.5%, surplus are water, pH6.6.
The fermentation culture process comprises: in initial 12 hours, the interval ventilation remains on the aerobic conditions fermentation, air flow 1: 1, and regulation and control fermentation dissolved oxygen 10%, mixing speed 180r/min, stirred 40 ℃ of temperature 2 minutes at 2 hours mixing chamber intervals; Slightly soluble oxygen and anaerobism cultivation stage: after 12 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation, stir at the interval, 4 hours mixing chamber intervals, stirred 40 ℃ of temperature 4 minutes.
Expand numerous cultivation bacterium liquid with 500 liters and join 7800kg in the solid fermentation substratum.Described solid fermentation substratum is milk-acid bacteria Pediococcus acidilactici1.5%, yeast Pichia pastoris1%, rice bran (48%), wheat bran (47.5%), beans cypress (1%), protein powder (1%) stir, use the film capping, every stirring in 24 hours, cultivated 66 hours.
Expand numerous bacterial classification according to liquid fermentation process in example 2.
Wherein, step 1), 2), yeast and milk-acid bacteria substratum used are proteolytic enzyme peptone (protease peptone) 1g, beef extract (beef extract) 1g, glucose (glucose) 2g, sodium acetate (sodium acetate) 3g, and Trisodium Citrate (ammoni acitrate) 2g.
The formula of substratum step 3) used is by mass percentage: Semen Maydis powder 1.0%, soyflour 2%, peptone 2.5%, yeast powder 2%, K
2HPO
41.5%, MgSO
40.1%, tween 80 1.5%, sodium-acetate 1.5%, surplus are water, pH6.8.
The fermentation culture process comprises: in initial 12 hours, the interval ventilation remains on the aerobic conditions fermentation, air flow 1: 1.2, and regulation and control fermentation dissolved oxygen 10%, mixing speed 180r/min, stirred 42 ℃ of temperature 2 minutes at 2 hours mixing chamber intervals; Slightly soluble oxygen and anaerobism cultivation stage: after 12 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation, stir at the interval, 4 hours mixing chamber intervals, stirred 42 ℃ of temperature 4 minutes.
Expand numerous cultivation bacterium liquid with 500 liters and join 8000kg in the solid fermentation substratum.Described solid fermentation substratum is milk-acid bacteria Pediococcus acidilactici1.5%, yeast Pichia pastoris1%, rice bran (43%), wheat bran (43%), beans cypress (6%), protein powder (5.5%) stir, use the film capping, every stirring in 24 hours, cultivated 72 hours.
Application examples
(1) this time be 30 days experimental period; cultivate flow process according to the solid fungicide of example 2-5 and be made into product; be JH1, JH2, JH3, JH4 to 4 kinds of product number consecutivelies; added up the number of viable of each microbial inoculum every 5 days; following diagram data shows that the JH1 microbial inoculum of example 2 correspondences is best to the absorption protection effect of microorganism, and number of viable remained on 6.5 * 10 in 30 days
6Cfu/g, survival rate reaches 85%, and the JH3 microbial inoculum effect of example 3 correspondences is the poorest, and number of viable dropped to 4.2 * 10 in 30 days
5Cfu/g, survival rate drops to 6.5%.The sequence that draws thus four kinds of cultural methods is JH1＞JH2＞JH4＞JH3, and best training method is example 2.
4 kinds of composite fungus agents of table 1 are processed the number of viable of 30 days
(2) this time be 14 days experimental period, in the aerobic kitchen garbage treater of 1kg treatment capacity, adds continuously the changing food waste of 540g at least, the highest 1746g that adds of Dan Tian every day.In the middle of added the rubbish such as chicken bone, Fishbone, dish class, rice, meat.Its method is: described bacterial classification is through liquid fermenting propagation, and field planting makes solid fungicide JH1 in solid filler, and this microbial inoculum is in being added to kitchen garbage treater with 1% addition.From experiment effect, carrier microbial inoculum quality does not increase, and the non-stimulated smell of decomposition course produces, and it is more than 96% that whole changing food waste is cleared up rate.Contrast before and after adding from changing food waste, after 4 hours, the changing food waste base conditioning is complete, without obviously residual.The material decomposition such as bone are slower, and decompose at least by the above time of two weeks for needs.In this section treating processes, water ratio tends towards stability, and in the handler carrier, temperature is 50 degrees centigrade of left and right, and microorganism growth is good, without the anaerobically fermenting phenomenon.
Table 2 is processed the rubbish total Amount Monitoring result after 14 days
| Experimental period (my god) | Gross weight (g) before processing | Add rubbish total amount (g) | Gross weight (g) after processing |
| 14 | 7000 | 8572 | 7337 |
(3) this time be 28 days experimental period, and experimental installation is the aerobic kitchen garbage treater of day output 50kg.Its method is: described bacterial classification is through liquid fermenting propagation, and field planting makes solid fungicide JH1 in solid filler, and this microbial inoculum is in being added to kitchen garbage treater with 1% addition.It is 450kg that filler adds total amount, 28 days process the changing food waste total amount is 1390kg, last surplus total amount is 380kg, changing food waste is in complete treated state, because a part self filler is also utilized by microbial consumption, and discharge with the heat energy form, cause the surplus total amount lower than adding the filler total amount.
Table 3 is processed the rubbish total Amount Monitoring result after 28 days
| Time (my god) | Add quantity of refuse (kg) | Add filler (kg) | Unspent amount (kg) | Decomposition efficiency |
| 1 | 20 | 400 | ||
| 2 | 20 | |||
| 3 | 20 |
[0059]
| 4 | 30 | 50 | ||
| 5～26 | 50 | |||
| 27 | 100 | |||
| 28 | 100 | |||
| Total amount | 1390 | 450 | 380 | 100％ |
(4) be 60 days this experimental period, and experimental installation is the aerobic kitchen garbage treater of day output 1kg.Its method is: make microbial inoculum according to table 4 method, in being added to kitchen garbage treater with 1% addition respectively, the filler gross weight is 7kg with microbial inoculum.Monitor its changes in weight situation every day.As seen from Figure 1, rubbish accumulative total was added 15kg in 60 days, and the composite fungus agent processing power of HBS-1 is the strongest, and the rubbish semi-invariant is minimum, and after 60 days, weight is 6.2kg, compares changing food waste and adds total amount, and rubbish decomposes fully.Yet, only adding the microbial inoculum of milk-acid bacteria, treatment effect is slightly poor, and after 60 days, weight is 8.7kg, and processing rate is 89%.Only add saccharomycetic treatment group effect the poorest, after 60 days, weight is 10.5kg, and processing rate is 77%.Thus, can draw, add composite fungus agent and be better than the treatment effect that only adds single bacterium.
The matching method of table 4 different strain addition
(5) be 30 days this experimental period, and experimental installation is the aerobic kitchen garbage treater of day output 1kg, and an other strain Angel Yeast is buied from the supermarket.Its method is: make microbial inoculum according to table 5 method, in being added to kitchen garbage treater with 1% addition respectively, the filler gross weight is 7kg with microbial inoculum.Monitor its changes in weight situation every day.As seen from Figure 2, rubbish accumulative total was added 10kg in 30 days, added the microbial inoculum of yeast (Pichia pastoris), and treatment effect is relatively obvious, and after 30 days, gross weight is 9.8kg, and processing rate is 72%.Add the treatment group effect of Angel Yeast relatively relatively poor, after 30 days, weight is 12.6kg, and processing rate is 44%.Thus, can draw, yeast treatment effect of the present invention is relatively obvious, the Angel Yeast that is better than buying on market.
The matching method of table 5 different strain addition
The above % unit all is weight percentage.
The above is only the preferred embodiment of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (3)
1. a changing food waste is subdued type yeast (Pichia pastoris) HBS-Hab, and its deposit number is CGMCC No.5960.
2. the application of yeast in decomposing changing food waste as claimed in claim 1.
3. the application of yeast in the composite fungus agent of preparation decomposition changing food waste as claimed in claim 1.
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| CN111715679B (en) * | 2020-07-21 | 2023-12-19 | 杭州洁洁环保科技有限公司 | Circulation dehydration biochemical treatment machine and control method thereof |
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| CN102391950A (en) * | 2011-11-17 | 2012-03-28 | 苏柯汉（潍坊）生物工程有限公司 | Food waste deodorization composite bacteria and preparation method thereof |
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| CN102174401A (en) * | 2011-01-30 | 2011-09-07 | 宁波大学 | Microorganism composite zymocyte agent as well as preparation method and application thereof |
| CN102391950A (en) * | 2011-11-17 | 2012-03-28 | 苏柯汉（潍坊）生物工程有限公司 | Food waste deodorization composite bacteria and preparation method thereof |
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Address after: 100089 No. 27 West Third Ring Road North, Beijing, Haidian District Patentee after: Institute of resources and environment, Beijing Academy of science and technology Address before: 100089 No. 27 West Third Ring Road North, Beijing, Haidian District Patentee before: ENVIRONMENTAL PROTECTION Research Institute OF LIGHT INDUSTRY |