CN110438179A - A method of γ-decalactone is prepared using kitchen waste oil - Google Patents

A method of γ-decalactone is prepared using kitchen waste oil Download PDF

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CN110438179A
CN110438179A CN201910489054.1A CN201910489054A CN110438179A CN 110438179 A CN110438179 A CN 110438179A CN 201910489054 A CN201910489054 A CN 201910489054A CN 110438179 A CN110438179 A CN 110438179A
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waste oil
rouge
kitchen waste
decalactone
gdl
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CN110438179B (en
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朱廷恒
王荣霞
汪琨
王渭霞
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Zhejiang University of Technology ZJUT
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    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/02—Oxygen as only ring hetero atoms
    • C12P17/04—Oxygen as only ring hetero atoms containing a five-membered hetero ring, e.g. griseofulvin, vitamin C

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Abstract

γ-decalactone method is prepared using kitchen waste oil the invention discloses a kind of, Yarrowia lipolytica suspension reconciliation rouge Candida bacteria suspension is mixed, it is seeded in the fermentation medium of the waste oil containing kitchen, 28~30 DEG C of cultures 20~for 24 hours, obtain the fermentation liquid containing Yarrowia lipolytica and Candida lipolytica；Fermentation liquid is seeded in the GDL conversion culture medium of ricinoleic acid containing 60-80g/L or castor oil with the inoculum concentration of volumetric concentration 10%, in 28 DEG C of shaken cultivation 72h, GDL conversion production is carried out, obtains γ-decalactone；For the present invention using kitchen waste grease as part carbon source, success culture yeasts bacterial strain simultaneously carries out GDL conversion production.The culture medium and mixed fungus fermentation method cost reduce, and turn waste into wealth, not only produce the GDL of high added value, but also carried out resource utilization to kitchen waste grease, have apparent economy, environmental benefit.

Description

A method of γ-decalactone is prepared using kitchen waste oil
(1) technical field
The present invention relates to a kind of using part carbon source in kitchen waste oil substitution complete medium, utilizes the ability of cutting grease Strong yeast Hybrid NC machine tool makes full use of grease, obtains containing the Yarrowia lipolytica compared with multi-biomass, recycles solution rouge Ye Shi Yeast conversion ricinoleic acid generates food-grade natural perfume material γ-decalactone.
(2) background technique
γ-decalactone (γ-decalactone, GDL) is a kind of lactone being naturally present in the fruit such as strawberry, peach Class fragrance, have lower fragrance threshold value, be internationally recognized safe food additives, be widely used in allotment coconut, strawberry, The fragrance such as peach are widely applied, market demand in margarine, ice cream, soft drink, candy, bakery product and seasoning It is very big.With the continuous growth to consumption such as ticbit, beverages, the demand of natural flavor additive is continuously increased.
Past, GDL are mainly extracted from natural biologicals such as plant, fruit or are synthesized using chemical method, with Rising of the consumer to natural prodcuts demand is had begun at present using biofermentation and transformation technology production GDL.Bioanalysis is raw The GDL of production is closer with natural plant composition, has obtained the attention of height.Microbial method is to produce safer, high biology to live The most promising method of perfume compound and its compound of property.The bioconversion of GDL be using fatty acid and aliphatic ester as Substrate carries out microbe conversion production using microorganism.Microbial strains have Yarrowia lipolytica (Yarrowia Lipolytica), saccharomyces cerevisiae (Saccharomyces cerevisiae) etc., wherein Yarrowia lipolytica is production GDL Main bacterial strain.
The amount of the kitchen waste grease in China is very huge, if not being pocessed utilization, will become " swill oil ", " gutter oil ", it is very harmful.Therefore, it develops this waste resource and realized valueization utilizes and is important solution.It utilizes Microorganism carries out conversion processing to grease is abandoned, and generating economic value high raw material, natural products etc. is a feasible approach.
The enzyme of the microorganism equal lipoid materials rich in that cut grease, can carry out biology to oil substances substrate Catalyzed conversion.The Ye Shi solution rouge yeast production GDL reported at present be all using complete medium, containing yeast extract, peptone, Glucose etc., industrialized production it is at high cost.If can be obtained using cheap lipid material as substrate carbon source culture Family name carries out GDL production after solving rouge yeast, then industrial production cost can be significantly reduced.Known Yarrowia lipolytica (and opinion Rouge Candida) a large amount of lipase and esterase can be secreted, it can decompose and be grown using lipid material, but solve rouge Ye Shi Yeast different strains inulinase-producing activity and GDL yield differ greatly, if carrying out ferment as medium component using kitchen waste oil Mother cultivates and converts production GDL, is a kind of both economically and environmentally beneficial method, is the approach for having very much production application potentiality, so far The present there is no relevant report.
The present invention utilizes solution rouge Ye Shi ferment by part carbon source and nutritional ingredient in addition kitchen waste oil substitutive medium Female (CICC 32187) and Candida lipolytica (CICC 31223) mixed fungus fermentation Synergistic degradation waste oil obtains solution rouge Ye Shi ferment GDL conversion production is carried out after certain biomass yeast based on mother again.
(3) summary of the invention
γ-decalactone method is prepared using kitchen waste oil it is an object of the present invention to provide a kind of, utilizes kitchen waste oil Instead of part carbon source in complete medium, reduce glucose content, reduce yeast extract content, makes full use of solution rouge Family name's yeast and Candida lipolytica mixed fungus fermentation obtain the Yarrowia lipolytica compared with multi-biomass and convert ricinoleic acid generation GDL。
The technical solution adopted by the present invention is that:
The present invention, which provides, a kind of prepares γ-decalactone method, the method using kitchen waste oil are as follows: (1) will solve rouge Ye Shi yeast bacteria suspension conciliates the mixing of rouge Candida bacteria suspension, obtains mixed bacteria liquid；By mixed bacteria liquid with volumetric concentration 10% Inoculum concentration be seeded in the fermentation medium of the waste oil containing kitchen, 28~30 DEG C culture 20~for 24 hours, obtain containing solution rouge Ye Shi ferment Female and Candida lipolytica fermentation liquid；The fermentation medium quality group becomes: glucose 1.25%, peptone 2%, yeast Powder 0.5%, kitchen waste oil 0.3~0.6% (preferably 0.6%), 1% Tween-80, solvent are deionized water, and pH value is natural； (2) fermentation liquid is seeded to the GDL conversion training of ricinoleic acid containing 60-80g/L or castor oil with the inoculum concentration of volumetric concentration 10% It supports in base, cultivates 72h in 28 DEG C of oscillations (200r/min), carry out GDL conversion production, conversion fluid is isolated and purified, obtained in γ-last of the ten Heavenly stems Ester；The GDL conversion culture medium quality composition are as follows: kitchen waste oil 0.6~4%, Tween-80 1%, KH2PO40.055%, Na2HPO40.031%, anhydrous MgSO40.016%, solvent is deionized water, and pH is natural.
Further, the Yarrowia lipolytica suspension conciliates rouge Candida bacteria suspension OD600Value is 0.1, described mixed Combined bacteria liquid is that 1:10 is mixed by volume by Yarrowia lipolytica suspension reconciliation rouge Candida bacteria suspension.
Further, the Yarrowia lipolytica is preferably Yarrowia lipolytica (Yarrowia lipolytica) CICC 32187, the Candida lipolytica is preferably Candida lipolytica (Candida lipolytica) CICC 31223.
Further, the Yarrowia lipolytica suspension is the preparation method comprises the following steps: (1) activation culture: Yarrowia lipolytica is connect Kind to YPD culture medium, 28 DEG C of cultures are activated for 2 days, obtain the single colonie of activation；The YPD culture medium quality composition: grape Sugar 2%, peptone 2%, yeast powder 1%, agar 2%, solvent are deionized water, and pH value is natural；
(2) seed culture: the single colonie (preferably 3~5) of picking activation is aseptically inoculated in equipped with 300mL kind In the 500mL conical flask of sub- culture medium, 28~30 DEG C, 150rpm shaking table culture 1 day obtain Yarrowia lipolytica seed liquor；With Sterile water adjusts seed liquor OD600Value is 1, as Yarrowia lipolytica suspension；The seed culture medium quality composition: grape Sugar 2%, peptone 2%, yeast powder 1%, solvent are deionized water, and pH value is natural.Candida lipolytica bacterium of the present invention is outstanding The preparation of liquid is the same as Yarrowia lipolytica suspension.
Further, kitchen waste oil of the present invention is prepared as follows: kitchen waste grease can be from meal The food waste waste oil that grease or other approach in the garbage collection process station of kitchen after water-oil separating obtain, can also voluntarily prepare. Specific preparation method takes humid heat treatment and centrifugal separation: kitchen garbage is removed bone, throwaway chopsticks, polybag and meal After the impurity such as towel paper, kitchen residue containing greasy is placed in the containers such as beaker, is placed in 80 DEG C of heating water bath 30min, It is centrifugated in 2500r/min, collecting upper layer waste oil is kitchen waste oil.
Further, γ-decalactone sample isolation and purification method in the conversion fluid are as follows: take fermentation liquid, 8000rpm/min from Supernatant is transferred to separatory funnel by heart 10min, and ethyl acetate is added in 1:1 by volume, and concussion is mixed, is stored at room temperature 10min, upper organic phase anhydrous Na2SO4It is dehydrated (0.2g/mL), GDL sample is obtained after filtering, can be used for vapor detection, such as Fruit needs to prepare a large amount of GDL, and supercritical carbon dioxide extraction method can be taken to carry out (the γ-of Su Chang supercritical carbon dioxide The Chinese spices and essence scientific seminar collection of thesis 2006:3 of decalactone extraction research .2006).
The present invention filters out high yield GDL respectively and the ability that reduces fat is strong by the screening test to different saccharomycete Yeast strain, the ability that wherein Candida lipolytica (CICC 31223) cuts grease is strong, Yarrowia lipolytica (CICC 32187) GDL high is produced, after further repeatedly cultivating domestication to the bacterial strain, improves GDL yield.
Compared with prior art, beneficial effect of the present invention is mainly reflected in:
1) existing GDL using microbe conversion method produce, fermentation medium quality group become complete medium (Braga A, Belo I.Production ofγ-decalactone by Yarrowia lipolytica:insights into experimental conditions and operating mode optimization[J].Journal of Chemical Technology&Biotechnology, 2015,90 (3): 559-565), consisting of: glucose 2%, albumen Peptone 2%, yeast powder 1%, the culture medium cost are high.The present invention utilizes kitchen waste oil substitution part glucose and yeast powder, training Support base composition are as follows: glucose 1.25%, peptone 2%, yeast powder 0.5%, kitchen waste oil 0.3~0.6%；Save production Cost.
2) Candida lipolytica (CICC 31223) and Yarrowia lipolytica (CICC 32187) Hybrid NC machine tool is utilized.
To sum up, for the present invention using kitchen waste grease as part carbon source, success culture yeasts bacterial strain simultaneously carries out GDL conversion Production.The culture medium and mixed fungus fermentation method cost reduce, and turn waste into wealth, not only produce the GDL of high added value, but also to kitchen Waste grease has carried out resource utilization, has apparent economy, environmental benefit.Microbe transformation method produces natural GDL and faces Main problem be reduce industrialized production cost.This can by develop cheap microbial fermentation raw material, strain transformation, It improves fermentation and conversion process etc. to carry out, there is very big industrial applications prospect.
(4) Detailed description of the invention
The ability of the decomposition Ester of Fig. 1 Yarrowia lipolytica and Candida lipolytica, Y.lipolytica solve rouge Family name's yeast CICC32187；C lipolytica, Candida lipolytica CICC31223.
Fig. 2 Yarrowia lipolytica and Candida lipolytica produce GDL measurement.
(5) specific embodiment
The present invention is described further combined with specific embodiments below, but protection scope of the present invention is not limited in This:
Yarrowia lipolytica (Yarrowia lipolytica) CICC 32187 of the present invention is purchased from Chinese industrial microorganism Culture Collection Center, bacterial strain deposit number CICC 32187.Candida lipolytica (Candida lipolytica) CICC31223 is purchased from Chinese industrial Culture Collection, bacterial strain deposit number CICC 31223.
Kitchen waste oil is prepared as follows in the embodiment of the present invention: kitchen waste oil derives from rubbish from cooking, takes Humid heat treatment and centrifugal separation preparation: kitchen garbage is removed into the impurity such as bone, throwaway chopsticks, polybag and napkin paper Afterwards, kitchen residue containing greasy is placed in beaker, is placed in 80 DEG C of heating water bath 30min, in 2500r/min centrifugation point From collecting upper layer waste oil is kitchen waste oil.
Embodiment 1 is to two plants of yeast degradation greases and produces GDL measurement
1, the ability of different yeast strain degradation esters is measured.
Ester and lactone generate hydroxamic acid with azanol reaction under alkaline condition, in acid condition hydroxamic acid and and Fe The aubergine complex generated is complexed, in OD506Nm has absorption.
By the screening verification to various bacterial strains, Yarrowia lipolytica (Yarrowia is finally picked them separately Lipolytica) CICC 32187 and 31223 single colonie of Candida lipolytica (Candida lipolytica) CICC are inoculated in YPD fluid nutrient medium 5mL, 28 DEG C of oscillation (200r/min) overnight incubations；It is forwarded to respectively with the inoculum concentration of volumetric concentration 10% In YPD fluid nutrient medium 30mL, 28 DEG C of oscillations (200r/min) cultivate 12h, 6 000r/min, are centrifuged 3min, and bacterial sediment is used Sorbierite buffer washs 2 times, and cell concentration is adjusted to OD600The bacteria suspension that nm value is 1, is connect with the bacterium amount that connects of 8% (v/v) Kind is into the conversion culture medium of the castor oil containing 60g/L of 60mL/500mL.28 DEG C, 200r/min shaken cultivation, take every 12h 500 μ L fermentation liquids, 6 000r/min are centrifuged 3min, as to be tested after taking 1mL supernatant that deionized water is used to be diluted to 1.5mL Sample surveys ester content, and bacterial sediment is for surveying biomass.
Ester content measurement: the hydroxylamine hydrochloride ethyl alcohol of 1mL 0.5mol/L is added in test tube, then plus 300 μ L 6mol/L Sodium hydrate aqueous solution, be added 1.5mL sample to be tested；The aqueous hydrochloric acid solution of 500 μ L 4mol/L is added in acutely oscillation 100s； Become to solution and clarify, continuously add the ferric chloride in aqueous solution colour developing of 150 μ L 5% (w/v), microplate reader measures OD506Nm, every group 3 A Duplicate Samples, the result is shown in Figure 1.Measurement result shows, Candida lipolytica CICC31223 treated total ester content is lower than solution rouge Ye Shi yeast CICC32187 illustrates that the ability of Candida lipolytica CICC31223 degradation fat is strong.
The biomass after two saccharomycete culture 84h is close as the result is shown for biomass estimation, according to the amount folding with culture medium It calculates, the biomass of Yarrowia lipolytica CICC32187 is 52.1g/L, and the biomass of Candida lipolytica CICC31223 is 51.8g/L。
The YPD fluid nutrient medium quality composition: glucose 2%, peptone 2%, yeast powder 1%, solvent are deionization Water, pH value are natural.Culture medium sterilizes 20min at 121 DEG C.
The conversion culture medium (g/L): Tween-80 4, KH2PO40.55, Na2HPO40.31, anhydrous MgSO40.16, Solvent is deionized water, and pH is natural.
2, different yeast strains are measured and produces GDL ability.
Pick them separately Yarrowia lipolytica (Yarrowia lipolytica) CICC 32187 and Candida lipolytica 31223 single colonie of (Candida lipolytica) CICC is inoculated in YPD fluid nutrient medium 5mL, 28 DEG C of oscillations (200r/min) Overnight incubation；It is forwarded in 30mL YPD fluid nutrient medium with the inoculum concentration of volumetric concentration 5%, 28 DEG C of oscillation (200r/min) trainings 12h is supported, thalline were collected by centrifugation by 6 000r/min, 3min, washs 2 thallus with sorbierite buffer, cell concentration is adjusted to OD600The bacteria suspension that nm value is 1, respectively with the ricinoleic acid containing 60g/L for connecing bacterium amount and being seeded to 30mL/250mL of 8% (v/v) It converts in culture medium, 28 DEG C of oscillations (200r/min) cultivate 72h, obtain conversion fluid, measure conversion fluid using high resolution gas chromatography The yield of middle γ-decalactone.
Sample treatment: taking the conversion fluid of 200 μ L, mixes with the peach aldehyde aqueous solution of 200 μ L 1g/L and 500 μ L ethyl acetate, Oscillation mixes, and 8000rpm is centrifuged 1min, and 200 μ L supernatants are transferred to the centrifuge tube of new 1.5mL, and 1 μ L sample introduction is accurately taken to survey GDL content.
High resolution gas chromatography testing conditions: HP-INNOWAX capillary chromatographic column: (30m × 0.32mm × 0.25 μm)；Column Temperature: 180 DEG C of holdings 10min, carrier gas (N2) flow velocity 4.3mL/min, 1 μ L of sample volume；280 DEG C of fid detector temperature；Split ratio: 10:1。
As a result see Fig. 2, testing result is shown, Yarrowia lipolytica CICC32187 produces GDL amount and is higher than Candida lipolytica CICC31223。
Embodiment 2 carries out biofermentation to ricinoleic acid using the culture medium Hybrid NC machine tool of the waste oil containing kitchen and produces γ- Decalactone
1, Yarrowia lipolytica (Yarrowia lipolytica) CICC 32187 actication of culture: is conciliate into rouge vacation silk ferment Mother (Candida lipolytica) CICC 31223 is seeded to YPD culture medium respectively, and 28 DEG C of cultures are activated for 2 days, respectively Obtain the single colonie of activation.
The YPD culture medium quality composition: glucose 2%, peptone 2%, yeast powder 1%, agar 2%, solvent are to go Ionized water, pH value are natural.Culture medium sterilizes 20min at 121 DEG C.
2, seed culture: 3~5 single colonies of saccharomycete of the above-mentioned activation of picking are aseptically inoculated in respectively to be equipped with In the 500mL conical flask of 300mL seed culture medium, 28~30 DEG C, 150rpm shaking table culture 1 day obtain solution rouge Ye Shi ferment respectively Female seed liquor and Candida lipolytica seed liquor.Dirt is not found by means inspections such as microscope detection, smell, color observations Dye.
Seed culture medium quality composition: glucose 2%, peptone 2%, yeast powder 1%, solvent are deionized water, pH value It is natural.
3, fermented and cultured: Yarrowia lipolytica seed liquor in step 2 is adjusted separately with aseptic deionized water first and conciliates rouge Candida seed liquor OD600Value is 1, obtains Yarrowia lipolytica suspension respectively and conciliates rouge Candida bacteria suspension.Then Candida lipolytica bacteria suspension and Yarrowia lipolytica suspension are mixed for 1:10 by volume, obtain mixed bacteria liquid.Press body Mixed bacteria liquid is inoculated into the 5L fermentor equipped with 3L fermentation medium into 28 DEG C of aerobic fementations by the inoculum concentration of product concentration 10% Culture 24 hours, ventilatory capacity control dissolved oxygen 8%, obtain fermentation liquid.
Fermentation medium quality composition: glucose 1.25%, peptone 2%, yeast powder 0.5%, kitchen waste oil 0.6%, 1% Tween-80, solvent is deionized water, and pH value is natural.
4, GDL conversion production: the fermentation liquid 3L access of step 3 is converted equipped with the 30L GDL for containing 60g/L ricinoleic acid In the 50L fermentor of culture medium, 72h is cultivated in 28 DEG C of oscillations (200r/min), carries out GDL conversion production.GDL converts culture medium Quality group becomes: kitchen waste oil 4%, Tween-80 1%, KH2PO40.055%, Na2HPO40.031%, anhydrous MgSO4 0.016%, solvent is deionized water, and pH is natural.
5, GDL assay: after conversion reaction, conversion fluid is directly taken to carry out GDL content using high resolution gas chromatography Measurement.As a result: every liter of fermentation output of fluid that conversion waste oil generates GDL is 0.33g/L.
γ-decalactone measurement method: it is measured using high resolution gas chromatography.
Sample treatment: taking the conversion fluid of 200 μ L, mixes with the peach aldehyde aqueous solution of 200 μ L 1g/L and 500 μ L ethyl acetate, Oscillation mixes, and 8000rpm is centrifuged 1min, and 200 μ L supernatants are transferred to the centrifuge tube of new 1.5mL, and 1 μ L sample introduction is accurately taken to survey GDL content.
High resolution gas chromatography testing conditions: HP-INNOWAX capillary chromatographic column: (30m × 0.32mm × 0.25 μm)；Column Temperature: 180 DEG C of holdings 10min, carrier gas (N2) flow velocity 4.3mL/min, 1 μ L of sample volume；280 DEG C of fid detector temperature；Split ratio: 10:1。
Embodiment 3 carries out fermenting and producing γ-decalactone to castor oil using the culture medium Hybrid NC machine tool of the waste oil containing kitchen
Ricinoleic acid in 2 step 4 of embodiment is changed to 80g/L castor oil, GDL converts kitchen waste oil matter in culture medium Amount concentration is changed to 3%, other operations are with embodiment 1, as a result: every liter of fermentation output of fluid that conversion waste oil generates GDL is 0.15g/L。

Claims (6)

1. a kind of prepare γ-decalactone method using kitchen waste oil, it is characterised in that the method are as follows: (1) rouge will be solved Family name's yeast bacteria suspension conciliates the mixing of rouge Candida bacteria suspension, obtains mixed bacteria liquid；By mixed bacteria liquid with volumetric concentration 10% Inoculum concentration is seeded in the fermentation medium of the waste oil containing kitchen, and 28~30 DEG C of cultures 20~for 24 hours, obtain fermentation liquid；The hair Ferment culture medium quality composition are as follows: glucose 1.25%, peptone 2%, yeast powder 0.5%, kitchen waste oil 0.3~0.6%, 1% Tween-80, solvent are deionized water, and pH value is natural；(2) fermentation liquid is seeded to the inoculum concentration of volumetric concentration 10% and is contained The GDL of 60-80g/L ricinoleic acid or castor oil is converted in culture medium, carries out γ-last of the ten Heavenly stems in 28 DEG C, 200r/min shaken cultivation 72h Lactone conversion, conversion fluid isolate and purify, and obtain γ-decalactone；The GDL conversion culture medium quality composition are as follows: kitchen waste oil 0.6~4%, 1% Tween-80, KH2PO40.055%, Na2HPO40.031%, anhydrous MgSO40.016%, solvent be go from Sub- water, pH are natural.
2. preparing γ-decalactone method using kitchen waste oil as described in claim 1, it is characterised in that the solution rouge Ye Shi Yeast bacteria suspension conciliates rouge Candida bacteria suspension OD600Value is 0.1.
3. preparing γ-decalactone method using kitchen waste oil as claimed in claim 2, it is characterised in that the solution rouge Ye Shi Yeast bacteria suspension conciliates rouge Candida bacteria suspension, and 1:10 is mixed by volume.
4. preparing γ-decalactone method using kitchen waste oil as described in claim 1, it is characterised in that the solution rouge Ye Shi Yeast is Yarrowia lipolytica (Yarrowia lipolytica) CICC 32187, and the Candida lipolytica is solution rouge vacation silk Yeast (Candida lipolytica) CICC 31223.
5. preparing γ-decalactone method using kitchen waste oil as described in claim 1, it is characterised in that the solution rouge Ye Shi Yeast bacteria suspension is the preparation method comprises the following steps: (1) activation culture: Yarrowia lipolytica is seeded to YPD culture medium, 28 DEG C of cultures 2 days into Row activation, obtains the single colonie of activation；The YPD culture medium quality composition: glucose 2%, peptone 2%, yeast powder 1%, Agar 2%, solvent are deionized water, and pH value is natural；
(2) seed culture: the single colonie of picking activation is aseptically inoculated in seed culture medium, 28~30 DEG C, 150rpm shaking table culture 1 day, obtain Yarrowia lipolytica seed liquor；Seed liquor OD is adjusted with aseptic deionized water600Value is 1, i.e., For Yarrowia lipolytica suspension；The seed culture medium quality composition: glucose 2%, peptone 2%, yeast powder 1% are molten Agent is deionized water, and pH value is natural；Preparation of the preparation of the Candida lipolytica bacteria suspension with Yarrowia lipolytica suspension.
6. preparing γ-decalactone method using kitchen waste oil as described in claim 1, it is characterised in that the kitchen waste oil Rouge is prepared as follows: after kitchen garbage is removed bone, throwaway chopsticks, polybag and napkin paper impurity, will contain oil The kitchen residue of rouge is centrifugated, collecting upper layer waste oil is kitchen in 80 DEG C of heating water bath 30min in 2500r/min Waste oil.
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刘莎等: "耶氏解脂假丝酵母（Yarrowia Lipolytica）发酵生产γ-癸内酯研究进展", 《粮食与油脂》 *
吴兰等: "解脂耶氏酵母处理含油废水的工艺条件", 《南昌大学学报(理科版)》 *
田凤蓉等: "酵母菌产乳化剂能力及其对含油废水降解性能的影响研究", 《环境工程学报》 *

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* Cited by examiner, † Cited by third party
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CN111394287A (en) * 2020-05-18 2020-07-10 中铁环境科技工程有限公司 Preparation method of degrading microbial inoculum for kitchen waste treatment
CN113186234A (en) * 2021-03-25 2021-07-30 安徽华业香料股份有限公司 Gamma-decalactone prepared based on biotransformation and preparation method thereof

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