CN105779551A

CN105779551A - Method for producing carotenoid through fermentation

Info

Publication number: CN105779551A
Application number: CN201610207683.7A
Authority: CN
Inventors: 李建树
Original assignee: BEIJING KEHUITONG WISDOM TECHNOLOGY Co Ltd
Current assignee: BEIJING KEHUITONG WISDOM TECHNOLOGY Co Ltd
Priority date: 2016-04-05
Filing date: 2016-04-05
Publication date: 2016-07-20

Abstract

The invention discloses a method for producing carotenoid through fermentation. The method comprises the following steps: by taking yellow peach cannery trimmings as a main raw material, carrying out microwave fixation, frozen-breaking and biological enzymolysis to obtain fruit juice with relatively high carotenoid content, adjusting components to obtain a fermentation medium, inoculating oceanic red yeast producing the carotenoid with high yield into the fermentation medium, carrying out fractional inoculation and temperature shift fermentation so that maximization of proliferation of oceanic red yeast is realized, carrying out ultrahigh pressure homogenization, vacuum centrifugation and demulsification separation on fermentation broth to obtain emulsion, and extracting, separating and purifying the emulsion to obtain the carotenoid finished product, wherein the cell biomass of the fermentation broth is 38-52g/L, and the yield of the carotenoid is 47-61mg/L. The preparation method is simple, short in period, high in efficiency, energy-saving and environment-friendly, the whole course adopts a low temperature biologic processing technology, the production cost is reduced, the hidden environmental pollution danger is eliminated, and a solid foundation is laid for sustainable development of the technical field of deep processing of yellow peaches, so that the method disclosed by the invention has positive economic and social development significance.

Description

A kind of method of fermenting and producing carotenoid

Technical field

The present invention relates to the preparation of carotenoid, a kind of method being specifically related to fermenting and producing carotenoid.

Background technology

Carotenoid is the general name of one group of fat-soluble pigment with non-oxidizability, and it is important as the class in humans and animals body Bioactive substance, has the important function such as enhancing human body immunity power, radioprotective, antitumor and treatment photosensitive diseases, at food The industries such as product, cosmetics, medicine and feedstuff have a wide range of applications.At present, carotenoid by FAO, the European Community and The international organizations such as WHO regard as A class nutrition pigment, and in recent years, around carotenoid industrialization development the most progressively Formed.

The industrialized preparing process of carotenoid can use chemical synthesis or plant extraction method.Chemosynthesis carotenoid Not only technical sophistication, and it uses as food additive and can affect the taste of food, reduces the ratio of absorption of human body, simultaneously There is also certain side effect.Therefore, the psychology advocating pollution-free food along with people strengthens day by day, the most more and more Research direction is turned to the development and utilization of natural pigment by scholar.Owing to extracting the conventional preparation techniques of carotenoid from plant Having higher cost and Operating Complexity, therefore, the industrialized production of carotenoid receives huge restriction.Utilize micro- Biofermentation produces carotenoid, the most with low cost, and technique is simple, and ensure that high-quality and the stable yields of product, Therefore, utilizing micro-organisms carotenoid is the main path that living resources type carotenoid is originated.

At present, the microbial strains that can produce carotenoid reported is mainly trispore Bruce mould and Rhodothece glutinis, the former Already at pilot scale and commercial application stage, the latter Ze Shang is in the laboratory lab scale stage.Although trispore Bruce mould pigment Yield is high, but culture process is complicated, and the production cycle is long, and Rhodothece glutinis produces carotenoid and then has nutritional requirement simply, Cultivation cycle is short, and thalline contains the advantage that rich in protein, aminoacid and vitamin etc. can comprehensively utilize.

Chinese patent CN105039484 A discloses one and utilizes ocean rhodotorula High Yield of Carotenoid and copper fermentation culture side Method, includes the following: described ocean rhodotorula WYN1 fermentation culture in cupric culture medium and efficiently produces carotenoid, fermentation Culture medium consists of glucose 1%, enzymolysis Semen Maydis powder 1-3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150-200mg/L, sodium chloride 20%, remaining is water；Initial pH is 8, inoculates 3-8%, shaking table Rotating speed is 50-80r/min, cultivates for 24-28 DEG C and adjusts temperature after 10-15 hour and be 16-20 DEG C and carry out low temperature static gas wave refrigerator, stops Only stirring, pH is adjusted to 3-5, keeps 2-4 hour；Stairstepping is warmed up to 24-30 DEG C afterwards, and pH is adjusted to 4.6-5.5, Addition according to 2.5% adds peptone, and the amount according to 2% adds yeast powder；Shaking speed is 50-100r/min, continues Supervention ferment 10-15h.

Chinese patent CN 104130952 A discloses a strain rhodotorula mucilaginosa and in fermenting and producing carotenoid and oils and fats Application.Technical scheme main points are: a Rhodotorula mucilaginose strain Rhodotorulamucilaginosa, in being preserved in State's Microbiological Culture Collection administration committee common micro-organisms center, CGMCCNo.8926.The rhodotorula mucilaginosa that the present invention provides can be used In fermenting and producing carotenoid and oils and fats.The present invention utilizes Rhodothece glutinis produce carotenoid and produce oils and fats simultaneously, and right Produce bacterial strain to carry out taking turns ARTP (atmospheric pressure at room plasma) mutation, it is thus achieved that the mutagenic strain that yield is higher, to class trailing plants recklessly more The industrialized production of Bu Su and oils and fats is provided with good application prospect and economic benefit.

Some antibacterials also can produce carotenoid: Chinese patent CN 104805168 A discloses one and utilizes photosynthetic bacteria micro- Aerobe fermentation quickly produces the method for carotenoid: by photosynthetic bacteria (Rhodobacter sphaeroides) inoculation of activation To MMS culture medium, aerobic cultivation to OD600 is 0.6-0.8, and then regulation condition of culture is to micro-oxygen, induces carotenoids Element rapid, high volume synthesis, then carries out extracting, purification, and the time utilizing the inventive method fermenting and producing carotenoid is short, only Need 36 hours, and thalline yield be up to 6g/L, the yield of carotenoid and productivity be respectively 8.287mg/L and 1911 μ g/g, have the ability of good removing DPPH free radical, and IC50 is about 8 μ g/mL.

Chinese patent CN 102732049 B discloses a kind of method that carotenoid is prepared in extraction from microbial cells.The party After method is microbial cells fermentation ends, fermentation liquid and thalline isolated wet thallus, then imported in cavity charge containers by logistics pipeline Carry out high steam explosion breaking cellular wall；Wet thallus dehydration after cell breakage, obtains the filter cake that bacterial chip is formed；Every kilogram of filter cake Add organic solvent 10～25 liters, 30～55 DEG C of stirring and leaching 20～50 minutes；Filter, obtain containing carotenoid oils and fats Organic solvent extracting solution；Extracting solution is concentrated in vacuo under the conditions of 40～60 DEG C, reclaims organic solvent, obtains carotenoid Oils and fats, single-trial extraction yield reaches more than 90%.This method saves the dehydrate in traditional handicraft and thalline pulverizing process, work Process flow is short, single-trial extraction yield high and the used time is short, organic solvent consumption reduces, and energy consumption and other costs significantly reduce.Whole Process is simple, quality controllable, is suitable for industrialized production application.

Patent disclosed above whether which kind of microorganism, its fermentation is prepared the raw material of carotenoid and is conventional medium dispensing Composition, relatively costly, and class Hu Luosu acquisition rate yield and productivity the most relatively low, be not suitable for current low-carbon (LC) produce environmental requirement.

Chinese patent CN 102286594 A discloses a kind of side utilizing high microsteping agricultural byproducts fermenting and producing carotenoid Method, is high microsteping agricultural byproducts raw material pulverizing to be processed, with adjuvant that is 0～6.0% ammonium sulfate and/or 0～3.0% phosphoric acid Potassium dihydrogen and/or 0～1.0% calcium chloride and/or 0～1.0% moisten water steaming after magnesium sulfate mix homogeneously；Steaming is cold after completing But 20～40 DEG C, sturdy vein born of the same parents bacterium, illumination cultivation 2～10d at 20～40 DEG C are connect；By cultured culture medium or collection spore Dried acid system breaking cellular wall, then extracts carotenoid by acetone or ethyl acetate；Carried pigment is separated, makes after purification Become carotenoid finished product.This method has process easy operation control, and production cost is low, and fermentation period is short, and carotenoid produces Amount advantages of higher.Patent fermentation period disclosed above is longer, and production cost is high, and yield and the productivity of carotenoid are still paid no attention to Think.

The nutrition of yellow peach is the abundantest, and its Major Nutrient composition has: the fibre that abundant vitamin C and substantial amounts of needed by human body are wanted Dimension element, carotene, lycoxanthin, red pigment and various trace elements.If selenium, zinc equal size are obviously higher than other common Fructus Persicae Son, possibly together with the composition such as malic acid, citric acid.Often eat yellow peach and the heat maintaining brain function can not only be provided, it is also possible to regulation Lipid metabolism in health, eats two every day and may only play relieving constipation, blood sugar lowering, blood fat, free radical resisting, dispel black speck, delay The effects such as old and feeble, raising immunologic function, also can promote appetite, can be rated as the Fructus Persicae of health fruit, health preserving.The most tired people, Pollute the people of environmental work, the people having a passion for one's pipe, be engaged in strenuous exercise and the people of highly intensive labour, Long-term taking medicine people the suitableeest Close and often eat yellow peach.

Due to yellow peach pole not shelf-stable, currently mainly based on canned food deep processing, the leftover bits and pieces that canned food produces, such as the peel reamed, The pit cut out and cull fruit etc., account for about the 30% of raw material weight, and major part rots, or is thrown away, macerates into fertilizer, some heaps Amass outside factory, cause public hazards, pollute environment and wastage of material is very big, add production cost simultaneously.In general, every hundred Gram yellow peach sarcocarp is containing carotenoid about between 1-10 milligram, and after can processing, carotenoid storage rate average out to 40% is left The right side, its amount containing carotenoid number be also one of the index weighing Yellow-peach can quality.The most except improving technique Reserved category carotene outside, add carotene also be improve yellow peach quality important means.Yellow peach peel is yellow peach tank One of leftover bits and pieces of head, every hectogram peel contains reducing sugar 2.4 grams, total protein 0.037 gram, carotene 0.0085 gram, goes back Containing multiple abundant vitamin.There is the most comprehensive recycle value.

At present, yet there are no and prepare pertinent literature or the report of carotenoid with Yellow-peach can leftover bits and pieces for fermenting raw materials.

Summary of the invention

Solved by the invention technical problem is that overcomes existing fermentation to prepare the defect of class Hu Luosu, based on Yellow-peach can leftover bits and pieces Want raw material, obtain, through microwave color fixing, freezing crushing, biological enzymolysis, the fruit juice that carotenoid content is higher, after adjusting component To fermentation medium, wherein the ocean rhodotorula of inoculation high yield class Hu Luosu carry out inoculating by several times, temperature-variable fermentation so that ocean Rhodothece glutinis maximizes propagation, obtains emulsion after fermentation liquor extra high pressure homogenize, traditional vacuum, break milk separation, and emulsion is through carrying Take, separate, obtain finished product carotenoid after purification.

In order to achieve the above object, the present invention is by the following technical solutions:

A kind of method of fermenting and producing carotenoid, comprises the steps:

1) prepared by fruit juice: first put in microwave dryer Yellow-peach can leftover bits and pieces in power 3-5kW, thickness of feed layer 2- 4cm, 105-115 DEG C, dry 2-4min, be then immersed in 10-15min in the sericin peptide taken solution that mass percent is 10-14%, Pulverize immediately after-18--22 DEG C of freezing 20-40min, freezing thickness of feed layer 3-5cm, ground product particle diameter 0.3-0.5mm, It is subsequently added into the water of ground product quality 1-3 times, is 3.5-5.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 25- 35kV/cm, burst length 300-500 μ s, carry out high voltage pulse electric field processing 20-under the conditions of pulse frequency 200-300Hz 30min；Then under the conditions of power 150-300W, microwave irradiation and extraction 15-20min is carried out in room temperature, simultaneously at power 200- 300W, carries out ultrasonic assistant extraction under the conditions of frequency 30-40KHz；Add the mixed enzyme of extracting solution quality 1.5-2.5%, in 40-50 DEG C of enzymolysis 30-50min, filters, obtains fruit juice；

Further, described Yellow-peach can leftover bits and pieces is: any one or two kinds in yellow peach peels, inferior yellow peach sarcocarp are with arbitrarily Mass ratio mixes；

Further, described mixed enzyme is cellulase, protease, pectase, tannase 2-4:2-4:1-in mass ratio 3:1-3 uniformly mixes；

2) prepared by fermentation medium: to step 1) fruit juice prepared adds the sugarcane of the protective agent of its quality 10-20%, 1-3% The magnesium sulfate of sugar, the ammonium sulfate of 0.5-1.5%, the potassium dihydrogen phosphate of 2-4%, the calcium chloride of 0.5-1.5% and 0.5-1.5% must be sent out Ferment culture medium；

Further, described protective agent with containing antifreeze protein, significantly improve Rhodothece glutinis and resist hot and cold irritability, disease resistance It is raw material with the plant of immunity, prepares through high-pressure pulse electric extraction, ultrasonic assistant microwave extraction and compound enzyme enzymolysis , the cold-and-heat resistent stress ability of Rhodothece glutinis in sweat can be effectively improved, improve its multiplication capacity and Viable detection, simultaneously Activity extra dry red wine yeast powder can be improved at freezing dry process Viable detection；

Preferably, described protectant preparation method, comprise the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, the Radix Astragali, Radix Codonopsis, excessive The sheep leaves of pulse plants, Herba Houttuyniae are respectively washed, drain, and 8-10:5-7:4-6:3-5:2-4:1-3 in mass ratio uniformly mixes, and add mixing The pH value of quality of material 0.1-1 times is the lactic acid moistening 3-8h of 3.8-4.5, carries out immediately after-18--22 DEG C of freezing 1-2h Pulverize, freezing thickness of feed layer 3-5cm, ground product particle diameter 0.5-3mm, be subsequently added into the water of ground product quality 10-20 times, with breast Acid for adjusting pH value is 3.5-5.5, at room temperature in electric field intensity 25-35kV/cm, and burst length 300-500 μ s, pulse frequency High voltage pulse electric field processing 20-30min is carried out under the conditions of 200-300Hz；Then enter under the conditions of power 150-300W in room temperature Row microwave irradiation and extraction 15-20min, simultaneously at power 200-300W, carries out ultrasonic assistant and carries under the conditions of frequency 30-40KHz Take；Add the compound enzyme of extracting solution quality 1-2%, in 45-55 DEG C of enzymolysis 30-50min；Enzymolysis solution filters, filtrate concentrates, Low-temperature grinding to particle diameter is that 0.1-0.3mm i.e. obtains protective agent；

Described compound enzyme is cellulase, protease, amylase, pectase, tannase 3-5:2-4:1-3:1-in mass ratio 3:1-2 uniformly mixes；

Extract it is highly preferred that described microwave irradiation and extraction is batch (-type), i.e. microwave exposure 10s, be spaced 20s；

3) fermentation, homogenizing breaking cellular wall: to step 2) fermentation medium adds the active extra dry red wine yeast powder of its quality 0.2-0.4% Controlling temperature 30-34 DEG C, speed of agitator 50-100r/min, ferment at constant temperature 12-15h, then with the speed of 0.8-1.0 DEG C/min Rate is warming up to 40-42 DEG C, adds the active extra dry red wine yeast powder constant temperature static fermentation 3-5h of fermentation medium quality 0.1-0.3%, After be cooled to 28-32 DEG C with the speed of 0.6-0.8 DEG C/min, continue static fermentation 15-20h, fermentation liquid successively through 40 mesh, 80 Mesh duplex strainer filter, filtrate through superhigh-voltage homogenizing machine in 2-4 DEG C, 140-160MPa homogenizing breaking cellular wall obtain homogenizing fluid；

Further, described activity extra dry red wine yeast powder is with ocean rhodotorula disclosed in Chinese patent CN105039484 A CCTCC No:M 2014592 is for set out what bacterium was prepared according to a conventional method；

Preferably, the protective agent used in activity extra dry red wine yeast powder preparation process is step 2) the middle protective agent prepared；

4) breakdown of emulsion, separation and Extraction: homogenizing fluid is put 6000-8000r/min traditional vacuum 10-15min in vacuum centrifuge, from Above lower isolated free oil, emulsion, hydrolyzed solution and solid residue, is 30-by the emulsion obtained control temperature 50 DEG C, add the biological demulsifying agent breakdown of emulsion 40-60min of emulsion quality 0.01-0.03%, in 2000-3000r/min after breakdown of emulsion Traditional vacuum 8-12min obtains free oil and emulsion again, and the free oil merging gained separately uses it for anything else, and gained emulsion is through carrying Take, separate, obtain finished product carotenoid after purification.

Further, described traditional vacuum condition is temperature 12-18 DEG C, vacuum-0.01--0.03MPa；

Further, described biological demulsifying agent is that glycolipid class, lipopeptid class, cell wall combine one or more in class biological demulsifying agent Combination；

Preferably, the quality group of described biological demulsifying agent becomes: glycolipid class: lipopeptid class: cell wall combines class=3-5:2-4:1-3；

Preferably, described glycolipid class biological demulsifying agent is one or both combinations in rhamnolipid, alkyl polyglucoside；

It is highly preferred that the quality group of described glycolipid class biological demulsifying agent becomes: rhamnolipid: alkyl polyglucoside=4-6:1-2；

Detection method and the assay method of carotenoid according to the cellular biomass disclosed in Chinese patent CN105039484 A The carrying out of carotenoid in cellular biomass in the fermentation liquid of above-mentioned preparation and emulsion is detected: cellular biomass is 38- 52g/L；Carotenoid output is 47-61mg/L.

Beneficial effect:

The present invention with Yellow-peach can leftover bits and pieces as raw material, initially with microwave drying while making leftover bits and pieces explosion puffing drying main It is enzyme denaturing, fixation, remains the natural colored of leftover bits and pieces and the content of carotenoid to greatest extent, after microwave color fixing Leftover bits and pieces natural colored is highly stable in the follow-up course of processing, will not variable color, fade, carotenoid content is the most steady Fixed, lose extremely low；In the aqueous solution containing sericin peptide taken, soak rehydration, leftover bits and pieces that is chilled and that cause can be reduced to greatest extent The loss of material active substance, improves the extraction ratio of leftover bits and pieces effective ingredient, is also follow-up alternating temperature activity extra dry red wine culture propagation simultaneously Establish solid foundation；The extract at low temperature technology such as high-pressure pulse electric, microwave, ultrasonic, biological enzymolysis are organically combined, can be Retain to limits bioactive ingredients content and the nutritional labeling such as class Hu Luosu of heel, effectively prevent course of processing miscellaneous bacteria dirty Dye；Science compounds protective agent in the fermentation medium, is remarkably improved ocean rhodotorula and resists hot and cold irritability, disease resistance And immunity so that it is fast activating, rejuvenation, it is achieved Rhodothece glutinis quality and quantity maximizes, and then realizes class Hu Luosu High yield；With the ocean rhodotorula of High Yield of Carotenoid as leaven, use temperature-variable fermentation and by several times vaccination ways can have Effect shortens lag phase and period of decline time, substantially prolongs exponential phase and stable phase, obtains ocean rhodotorula to greatest extent Propagation and the maximum accumulation of carotenoid, improve its multiplication capacity and Viable detection；Red by extra high pressure homogenize breaking cellular wall ocean Yeast cells, sporoderm-broken rate is high, and class Hu Luosu burst size is big；Traditional vacuum and biological demulsifying will be used to organically combine, significantly improve The extraction ratio of carotenoid, improves raw material availability, reduces extraction cost.Improving product quality and extraction ratio Add the kind of by-product after microbial grease etc. extracts, yield and added value simultaneously, reduce further carotenoid Extraction cost, has stopped the discharge of " three wastes ", has protected environment, is truly realized low-carbon (LC) and produces.

Detection method and the assay method of carotenoid according to the cellular biomass disclosed in Chinese patent CN105039484 A In cellular biomass in the fermentation liquid preparing the present invention and emulsion, the carrying out of carotenoid detects: cellular biomass is 38-52g/L；Carotenoid output is 47-61mg/L.

The preparation method of the present invention is simple, and the cycle is short, efficiency is high, save the energy, environmentally friendly, omnidistance uses low temperature, life Thing process technology, effectively prevent the loss of the heat-sensitive nutrition such as carotenoid, vitamin in leftover bits and pieces, improves class The extraction ratio of carotene and biological activity and nutrition content, improve yield and the quality of product, reduce and produce into This, a kind of method that simultaneously present invention also offers isolated microbial grease, there is preferable practicality and advance.

It should be noted that and the solution have the advantages that each synergistic summation of step technique feature, have between each step Certain inherent dependency, the simple superposition of the most single technical characteristic effect.

It can further be stated that the present invention prepares class Hu Luosu with Yellow-peach can leftover bits and pieces for raw material, the deep processing for yellow peach opens Article one, new shortcut, is effectively reduced the burden of processing enterprise, significantly reduces production cost, add value-added content of product, Eliminating environmental pollution hidden danger, solid foundation has been established in the sustainable development for the deep process technology field of yellow peach, has actively Economy and social development meaning.

Detailed description of the invention

The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention is this Method well known to skilled person.It addition, embodiment is interpreted as illustrative, and unrestricted the scope of the present invention, The spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, real without departing substantially from the present invention On the premise of matter and scope, the various changes or the change that carry out the material component in these embodiments and consumption fall within this Bright protection domain.

Embodiment 1

A kind of method utilizing yellow peach peels fermenting and producing carotenoid, comprises the steps:

1) prepared by fruit juice: first yellow peach peels is put in microwave dryer in power 4kW, thickness of feed layer 3cm, 110 DEG C, dry Dry 3min, is then immersed in 12min in the sericin peptide taken solution that mass percent is 12%, enters immediately after-20 DEG C of freezing 30min Row is pulverized, freezing thickness of feed layer 4cm, and ground product particle diameter 0.4mm is subsequently added into the water of ground product quality 2 times, regulates with lactic acid PH value is 4.5, at room temperature in electric field intensity 30kV/cm, and burst length 400 μ s, carry out height under the conditions of pulse frequency 250Hz Pressure impulse electric field processes 25min；Then under the conditions of power 200W, microwave irradiation and extraction 18min is carried out in room temperature, simultaneously in merit Rate 250W, carries out ultrasonic assistant extraction under the conditions of frequency 35KHz；Add the mixed enzyme of extracting solution quality 2.0%, in 45 DEG C Enzymolysis 40min, filters, obtains fruit juice；

Described mixed enzyme is that cellulase, protease, pectase, tannase 3:3:2:2 in mass ratio uniformly mixes；

2) prepared by fermentation medium: to step 1) fruit juice prepared adds the protective agent of its quality 15%, the sucrose of 2%, 1% The magnesium sulfate of ammonium sulfate, the potassium dihydrogen phosphate of 3%, the calcium chloride of 1% and 1% obtain fermentation medium；

Described protectant preparation method, comprises the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, the Radix Astragali, Radix Codonopsis, Herba Epimedii, fish Raw meat grass is respectively washed, drains, and 9:6:5:4:3:2 in mass ratio uniformly mixes, and adds the pH value of mixed material quality 0.5 times It is the lactic acid moistening 5h of 4.2, pulverizes immediately after-20 DEG C of freezing 1.5h, freezing thickness of feed layer 4cm, ground product particle diameter 2mm, is subsequently added into the water of ground product quality 15 times, is 4.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 30kV/cm, burst length 400 μ s, carry out high voltage pulse electric field processing 25min under the conditions of pulse frequency 250Hz；Then in room Temperature carries out microwave irradiation and extraction 18min, wherein, microwave exposure 10s under the conditions of power 200W, is spaced 20s；Simultaneously in merit Rate 250W, carries out ultrasonic assistant extraction under the conditions of frequency 35KHz；Add the compound enzyme of extracting solution quality 1.5%, in 50 DEG C Enzymolysis 40min；Enzymolysis solution filters, filtrate concentrates, low-temperature grinding to particle diameter is that 0.2mm i.e. obtains protective agent；

Described compound enzyme is that cellulase, protease, amylase, pectase, tannase 4:3:2:2:1.5 in mass ratio is uniform Mixing；

3) fermentation, homogenizing breaking cellular wall: to step 2) the active extra dry red wine yeast powder that adds its quality 0.3% in fermentation medium controls Temperature 32 DEG C, speed of agitator 80r/min, ferment at constant temperature 13h, then with the ramp of 0.9 DEG C/min to 41 DEG C, add The active extra dry red wine yeast powder constant temperature static fermentation 4h of fermentation medium quality 0.2%, is finally cooled to the speed of 0.7 DEG C/min 30 DEG C, continuing static fermentation 18h, fermentation liquid filters through 40 mesh, 80 mesh duplex strainers successively, and filtrate is through superhigh-voltage homogenizing machine In 3 DEG C, 150MPa homogenizing breaking cellular wall obtain homogenizing fluid；

Described activity extra dry red wine yeast powder is to prepare according to a conventional method with ocean rhodotorula CCTCC No:M 2014592 for the bacterium that sets out 's；

The protective agent used in described activity extra dry red wine yeast powder preparation process is step 2) the middle protective agent prepared；

4) breakdown of emulsion, separation and Extraction: homogenizing fluid is put 7000r/min traditional vacuum 12min in vacuum centrifuge, divides from top to bottom From obtaining free oil, emulsion, hydrolyzed solution and solid residue, it is 40 DEG C by the emulsion obtained control temperature, adds emulsus The biological demulsifying agent breakdown of emulsion 50min of liquid quality 0.02%, is dissociated in 2500r/min traditional vacuum 10min again after breakdown of emulsion Oil and emulsion, the free oil merging gained separately uses it for anything else, and gained emulsion is extracted, separate, obtain into category after purification recklessly Radix Raphani element；

Described traditional vacuum condition is temperature 15 DEG C, vacuum-0.02MPa；

The quality group of described biological demulsifying agent becomes: glycolipid class: lipopeptid class: cell wall combines class=4:3:2；

The quality group of described glycolipid class biological demulsifying agent becomes: rhamnolipid: alkyl polyglucoside=5:1.5.

In cellular biomass in fermentation liquid prepared by said method and emulsion, the carrying out of carotenoid detects: cellular biomass For 52g/L；Carotenoid output is 61mg/L.

Embodiment 2

A kind of method utilizing inferior yellow peach fruit pulp fermentation to produce carotenoid, comprises the steps:

1) prepared by fruit juice: first by inferior yellow peach sarcocarp section, puts in microwave dryer in power 3kW, thickness of feed layer 2cm, 105 DEG C, dry 2min, be then immersed in 10min in the sericin peptide taken solution that mass percent is 10%, in-18 DEG C of freezings Pulverize immediately after 20min, freezing thickness of feed layer 3cm, ground product particle diameter 0.3mm, be subsequently added into ground product quality 1 times Water, is 3.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 25kV/cm, and burst length 300 μ s, pulse frequency High voltage pulse electric field processing 20min is carried out under the conditions of 200Hz；Then under the conditions of power 150W, carry out microwave exposure in room temperature to carry Take 15min, simultaneously at power 200W, under the conditions of frequency 30KHz, carry out ultrasonic assistant extraction；Add extracting solution quality 1.5% Mixed enzyme, in 40 DEG C of enzymolysis 30min, filter, obtain fruit juice；

Described mixed enzyme is that cellulase, protease, pectase, tannase 2:2:1:1 in mass ratio uniformly mixes；

2) prepared by fermentation medium: to step 1) fruit juice prepared adds the protective agent of its quality 10%, the sucrose of 1%, The magnesium sulfate of the ammonium sulfate of 0.5%, the potassium dihydrogen phosphate of 2%, the calcium chloride of 0.5% and 0.5% obtains fermentation medium；

Described protectant preparation method, comprises the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, the Radix Astragali, Radix Codonopsis, Herba Epimedii, fish Raw meat grass is respectively washed, drains, and 8:5:4:3:2:1 in mass ratio uniformly mixes, and adds the pH value of mixed material quality 0.1 times It is the lactic acid moistening 3h of 3.8, pulverizes immediately after-18 DEG C of freezing 1h, freezing thickness of feed layer 3cm, ground product particle diameter 0.5mm, is subsequently added into the water of ground product quality 10 times, is 3.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 25kV/cm, burst length 300 μ s, carry out high voltage pulse electric field processing 20min under the conditions of pulse frequency 200Hz；Then in room Temperature carries out microwave irradiation and extraction 15min, wherein, microwave exposure 10s under the conditions of power 150W, is spaced 20s；Simultaneously in merit Rate 200W, carries out ultrasonic assistant extraction under the conditions of frequency 30KHz；Add the compound enzyme of extracting solution quality 1%, in 45 DEG C of enzymes Solve 30min；Enzymolysis solution filters, filtrate concentrates, low-temperature grinding to particle diameter is that 0.1mm i.e. obtains protective agent；

Described compound enzyme is that cellulase, protease, amylase, pectase, tannase 3:2:1:1:1 in mass ratio uniformly mixes Close；

3) fermentation, homogenizing breaking cellular wall: to step 2) the active extra dry red wine yeast powder that adds its quality 0.2% in fermentation medium controls Temperature 30 DEG C, speed of agitator 50r/min, ferment at constant temperature 12h, then with the ramp of 0.8 DEG C/min to 40 DEG C, add The active extra dry red wine yeast powder constant temperature static fermentation 3h of fermentation medium quality 0.1%, finally lowers the temperature with the speed of 0.6 DEG C/min To 28 DEG C, continuing static fermentation 15h, fermentation liquid filters through 40 mesh, 80 mesh duplex strainers successively, and filtrate is equal through supertension Matter machine in 2 DEG C, 140MPa homogenizing breaking cellular wall obtain homogenizing fluid；

4) breakdown of emulsion, separation and Extraction: homogenizing fluid is put 6000r/min traditional vacuum 10min in vacuum centrifuge, divides from top to bottom From obtaining free oil, emulsion, hydrolyzed solution and solid residue, it is 30 DEG C by the emulsion obtained control temperature, adds emulsus The biological demulsifying agent breakdown of emulsion 40min of liquid quality 0.01%, obtains free oil in 2000r/min traditional vacuum 8min again after breakdown of emulsion And emulsion, the free oil merging gained separately uses it for anything else, and gained emulsion is extracted, separate, obtain into category trailing plants recklessly after purification Bu Su；

Described traditional vacuum condition is temperature 12 DEG C, vacuum-0.01MPa；

The quality group of described biological demulsifying agent becomes: glycolipid class: lipopeptid class: cell wall combines class=3:2:1；

The quality group of described glycolipid class biological demulsifying agent becomes: rhamnolipid: alkyl polyglucoside=4:1.

In cellular biomass in fermentation liquid prepared by said method and emulsion, the carrying out of carotenoid detects: cellular biomass For 51g/L；Carotenoid output is 58mg/L.

Embodiment 3

A kind of method of fermenting and producing carotenoid, comprises the steps:

1) prepared by fruit juice: first by inferior for yellow peach sarcocarp section, then uniformly mixes with yellow peach peels 3:1 in mass ratio, puts into In power 5kW, thickness of feed layer 4cm, 115 DEG C, dry 4min in microwave dryer, being then immersed in mass percent is 14% Sericin peptide taken solution in 15min, pulverize immediately after-22 DEG C of freezing 40min, freezing thickness of feed layer 5cm, ground product grain Footpath 0.5mm, is subsequently added into the water of ground product quality 3 times, is 5.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 35kV/cm, burst length 500 μ s, carry out high voltage pulse electric field processing 30min under the conditions of pulse frequency 300Hz；Then in room Temperature carries out microwave irradiation and extraction 20min under the conditions of power 300W, simultaneously at power 300W, carries out under the conditions of frequency 40KHz Ultrasonic assistant extracts；Add the mixed enzyme of extracting solution quality 2.5%, in 50 DEG C of enzymolysis 50min, filter, obtain fruit juice；

Described mixed enzyme is that cellulase, protease, pectase, tannase 4:4:3:3 in mass ratio uniformly mixes；

2) prepared by fermentation medium: to step 1) fruit juice prepared adds the protective agent of its quality 20%, the sucrose of 3%, The magnesium sulfate of the ammonium sulfate of 1.5%, the potassium dihydrogen phosphate of 4%, the calcium chloride of 1.5% and 1.5% obtains fermentation medium；

Described protectant preparation method, comprises the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, the Radix Astragali, Radix Codonopsis, Herba Epimedii, fish Raw meat grass is respectively washed, drains, and 10:7:6:5:4:3 in mass ratio uniformly mixes, and the pH value adding mixed material quality 1 times is The lactic acid moistening 8h of 4.5, pulverizes after-22 DEG C of freezing 2h immediately, freezing thickness of feed layer 5cm, ground product particle diameter 3mm, It is subsequently added into the water of ground product quality 20 times, is 5.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 35kV/cm, Burst length 500 μ s, carries out high voltage pulse electric field processing 30min under the conditions of pulse frequency 300Hz；Then in room temperature at power Carry out microwave irradiation and extraction 20min, wherein, microwave exposure 10s under the conditions of 300W, be spaced 20s；Simultaneously at power 300W, Ultrasonic assistant extraction is carried out under the conditions of frequency 40KHz；Add the compound enzyme of extracting solution quality 2%, in 55 DEG C of enzymolysis 50min；Enzymolysis solution filters, filtrate concentrates, low-temperature grinding to particle diameter is that 0.3mm i.e. obtains protective agent；

Described compound enzyme is that cellulase, protease, amylase, pectase, tannase 5:4:3:3:2 in mass ratio uniformly mixes Close；

3) fermentation, homogenizing breaking cellular wall: to step 2) the active extra dry red wine yeast powder that adds its quality 0.4% in fermentation medium controls Temperature 34 DEG C, speed of agitator 100r/min, ferment at constant temperature 15h, then with the ramp of 1.0 DEG C/min to 42 DEG C, add Enter the active extra dry red wine yeast powder constant temperature static fermentation 5h of fermentation medium quality 0.3%, finally lower the temperature with the speed of 0.8 DEG C/min To 32 DEG C, continuing static fermentation 20h, fermentation liquid filters through 40 mesh, 80 mesh duplex strainers successively, and filtrate is equal through supertension Matter machine in 4 DEG C, 160MPa homogenizing breaking cellular wall obtain homogenizing fluid；

4) breakdown of emulsion, separation and Extraction: homogenizing fluid is put 8000r/min traditional vacuum 15min in vacuum centrifuge, divides from top to bottom From obtaining free oil, emulsion, hydrolyzed solution and solid residue, it is 50 DEG C by the emulsion obtained control temperature, adds emulsus The biological demulsifying agent breakdown of emulsion 60min of liquid quality 0.03%, is dissociated in 3000r/min traditional vacuum 12min again after breakdown of emulsion Oil and emulsion, the free oil merging gained separately uses it for anything else, and gained emulsion is extracted, separate, obtain into category after purification recklessly Radix Raphani element；

Described traditional vacuum condition is temperature 18 DEG C, vacuum-0.03MPa；

The quality group of described biological demulsifying agent becomes: glycolipid class: lipopeptid class: cell wall combines class=5:4:3；

The quality group of described glycolipid class biological demulsifying agent becomes: rhamnolipid: alkyl polyglucoside=6:2.

In cellular biomass in fermentation liquid prepared by said method and emulsion, the carrying out of carotenoid detects: cellular biomass For 45g/L；Carotenoid output is 52mg/L.

Embodiment 4

1) prepared by fruit juice: first yellow peach peels is put in microwave dryer in power 3kW, thickness of feed layer 4cm, 105 DEG C, dry Dry 4min, is then immersed in 15min in the sericin peptide taken solution that mass percent is 10%, enters immediately after-18 DEG C of freezing 40min Row is pulverized, freezing thickness of feed layer 3cm, and ground product particle diameter 0.5mm is subsequently added into the water of ground product quality 1 times, regulates with lactic acid PH value is 5.5, at room temperature in electric field intensity 25kV/cm, and burst length 500 μ s, carry out height under the conditions of pulse frequency 200Hz Pressure impulse electric field processes 30min；Then under the conditions of power 150W, microwave irradiation and extraction 20min is carried out in room temperature, simultaneously in merit Rate 200W, carries out ultrasonic assistant extraction under the conditions of frequency 40KHz；Add the mixed enzyme of extracting solution quality 1.5%, in 50 DEG C Enzymolysis 30min, filters, obtains fruit juice；

Described mixed enzyme is that cellulase, protease, pectase, tannase 2:4:1:3 in mass ratio uniformly mixes；

2) prepared by fermentation medium: to step 1) fruit juice prepared adds the protective agent of its quality 10%, the sucrose of 3%, The magnesium sulfate of the ammonium sulfate of 0.5%, the potassium dihydrogen phosphate of 4%, the calcium chloride of 0.5% and 1.5% obtains fermentation medium；

Described protectant preparation method, comprises the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, the Radix Astragali, Radix Codonopsis, Herba Epimedii, fish Raw meat grass is respectively washed, drains, and 8:7:4:5:2:3 in mass ratio uniformly mixes, and adds the pH value of mixed material quality 0.1 times It is the lactic acid moistening 3h of 4.5, pulverizes immediately after-22 DEG C of freezing 1h, freezing thickness of feed layer 3cm, ground product particle diameter 3mm, is subsequently added into the water of ground product quality 10 times, is 5.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 25kV/cm, burst length 500 μ s, carry out high voltage pulse electric field processing 30min under the conditions of pulse frequency 200Hz；Then in room Temperature carries out microwave irradiation and extraction 20min under the conditions of power 150W, simultaneously at power 200W, carries out under the conditions of frequency 40KHz Ultrasonic assistant extracts；Add the compound enzyme of extracting solution quality 1%, in 55 DEG C of enzymolysis 30min；Enzymolysis solution filters, filtrate is dense Contracting, low-temperature grinding to particle diameter are that 0.3mm i.e. obtains protective agent；

Described compound enzyme is that cellulase, protease, amylase, pectase, tannase 3:4:1:3:1 in mass ratio uniformly mixes Close；

3) fermentation, homogenizing breaking cellular wall: to step 2) the active extra dry red wine yeast powder that adds its quality 0.2% in fermentation medium controls Temperature 34 DEG C, speed of agitator 50r/min, ferment at constant temperature 15h, then with the ramp of 0.8 DEG C/min to 42 DEG C, add The active extra dry red wine yeast powder constant temperature static fermentation 5h of fermentation medium quality 0.1%, is finally cooled to the speed of 0.6 DEG C/min 28 DEG C, continuing static fermentation 20h, fermentation liquid filters through 40 mesh, 80 mesh duplex strainers successively, and filtrate is through superhigh-voltage homogenizing machine Homogenizing fluid is obtained in 2 DEG C of 140-160MPa homogenizing breaking cellular walls；

4) breakdown of emulsion, separation and Extraction: homogenizing fluid is put 6000r/min traditional vacuum 15min in vacuum centrifuge, divides from top to bottom From obtaining free oil, emulsion, hydrolyzed solution and solid residue, it is 30 DEG C by the emulsion obtained control temperature, adds emulsus The rhamnolipid breakdown of emulsion 40min of liquid quality 0.03%, after breakdown of emulsion in 3000r/min traditional vacuum 8min again obtain free oil and Emulsion, the free oil merging gained separately uses it for anything else, and gained emulsion is extracted, separate, obtain finished product carotenoids after purification Element.

In cellular biomass in fermentation liquid prepared by said method and emulsion, the carrying out of carotenoid detects: cellular biomass For 42g/L；Carotenoid output is 48mg/L.

Embodiment 5

1) prepared by fruit juice: first yellow peach peels is put in microwave dryer in power 5kW, thickness of feed layer 2cm, 115 DEG C, dry Dry 2min, is then immersed in 10min in the sericin peptide taken solution that mass percent is 14%, enters immediately after-22 DEG C of freezing 20min Row is pulverized, freezing thickness of feed layer 5cm, and ground product particle diameter 0.3mm is subsequently added into the water of ground product quality 3 times, regulates with lactic acid PH value is 3.5, at room temperature in electric field intensity 35kV/cm, and burst length 300 μ s, carry out height under the conditions of pulse frequency 300Hz Pressure impulse electric field processes 20min；Then under the conditions of power 300W, microwave irradiation and extraction 15min is carried out in room temperature, simultaneously in merit Rate 300W, carries out ultrasonic assistant extraction under the conditions of frequency 30KHz；Add the mixed enzyme of extracting solution quality 2.5%, in 40 DEG C Enzymolysis 50min, filters, obtains fruit juice；

Described mixed enzyme is that cellulase, protease, pectase, tannase 4:2:3:1 in mass ratio uniformly mixes；

2) prepared by fermentation medium: to step 1) fruit juice prepared adds the protective agent of its quality 20%, the sucrose of 1%, The magnesium sulfate of the ammonium sulfate of 1.5%, the potassium dihydrogen phosphate of 2%, the calcium chloride of 1.5% and 0.5% obtains fermentation medium；

Described protectant preparation method, comprises the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, the Radix Astragali, Radix Codonopsis, Herba Epimedii, fish Raw meat grass is respectively washed, drains, and 10:5:6:3:4:1 in mass ratio uniformly mixes, and the pH value adding mixed material quality 1 times is The lactic acid moistening 8h of 3.8, pulverizes after-18 DEG C of freezing 2h immediately, freezing thickness of feed layer 3cm, ground product particle diameter 3mm, It is subsequently added into the water of ground product quality 10 times, is 5.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 25kV/cm, Burst length 500 μ s, carries out high voltage pulse electric field processing 30min under the conditions of pulse frequency 200Hz；Then in room temperature at power Carry out microwave irradiation and extraction 20min under the conditions of 150W, simultaneously at power 200W, carry out ultrasound wave under the conditions of frequency 40KHz auxiliary Help extraction；Add the compound enzyme of extracting solution quality 1%, in 55 DEG C of enzymolysis 30min；Enzymolysis solution filters, filtrate concentrates, low temperature Being crushed to particle diameter is that 0.3mm i.e. obtains protective agent；

Described compound enzyme is that cellulase, protease, amylase, pectase, tannase 5:2:3:1:2 in mass ratio uniformly mixes Close；

3) fermentation, homogenizing breaking cellular wall: to step 2) the active extra dry red wine yeast powder that adds its quality 0.4% in fermentation medium controls Temperature 30 DEG C, speed of agitator 100r/min, ferment at constant temperature 12h, then with the ramp of 1.0 DEG C/min to 40 DEG C, add Enter the active extra dry red wine yeast powder constant temperature static fermentation 3h of fermentation medium quality 0.3%, finally lower the temperature with the speed of 0.8 DEG C/min To 28 DEG C, continuing static fermentation 20h, fermentation liquid filters through 40 mesh, 80 mesh duplex strainers successively, and filtrate is equal through supertension Matter machine in 4 DEG C, 140MPa homogenizing breaking cellular wall obtain homogenizing fluid；

4) breakdown of emulsion, separation and Extraction: homogenizing fluid is put 8000r/min traditional vacuum 10min in vacuum centrifuge, divides from top to bottom From obtaining free oil, emulsion, hydrolyzed solution and solid residue, it is 50 DEG C by the emulsion obtained control temperature, adds emulsus The cell wall of liquid quality 0.01% combines class biological demulsifying agent breakdown of emulsion 60min, in 2000r/min traditional vacuum again after breakdown of emulsion 12min obtains free oil and emulsion, and the free oil merging gained separately uses it for anything else, and gained emulsion is extracted, separate, purification After obtain finished product carotenoid.

In cellular biomass in fermentation liquid prepared by said method and emulsion, the carrying out of carotenoid detects: cellular biomass For 38g/L；Carotenoid output is 47mg/L.

Claims

1. the method for a fermenting and producing carotenoid, it is characterised in that comprise the steps:

Described mixed enzyme is that cellulase, protease, pectase, tannase 2-4:2-4:1-3:1-3 in mass ratio uniformly mixes Close；

2) prepared by fermentation medium: to step 1) fruit juice prepared adds the sugarcane of the protective agent of its quality 10-20%, 1-3% The magnesium sulfate of sugar, the ammonium sulfate of 0.5-1.5%, the potassium dihydrogen phosphate of 2-4%, the calcium chloride of 0.5-1.5% and 0.5-1.5% obtains Fermentation medium；

Described protectant preparation method, comprises the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, the Radix Astragali, Radix Codonopsis, Herba Epimedii, fish Raw meat grass is respectively washed, drains, and 8-10:5-7:4-6:3-5:2-4:1-3 in mass ratio uniformly mixes, and adds mixed material quality The pH value of 0.1-1 times is the lactic acid moistening 3-8h of 3.8-4.5, pulverizes immediately after-18--22 DEG C of freezing 1-2h, freezing Thickness of feed layer 3-5cm, ground product particle diameter 0.5-3mm, it is subsequently added into the water of ground product quality 10-20 times, with breast acid for adjusting pH Value is 3.5-5.5, at room temperature in electric field intensity 25-35kV/cm, burst length 300-500 μ s, pulse frequency 200-300Hz Under the conditions of carry out high voltage pulse electric field processing 20-30min；Then under the conditions of power 150-300W, microwave exposure is carried out in room temperature Extract 15-20min, simultaneously at power 200-300W, under the conditions of frequency 30-40KHz, carry out ultrasonic assistant extraction；Addition carries Take the compound enzyme of liquid quality 1-2%, in 45-55 DEG C of enzymolysis 30-50min；Enzymolysis solution filtration, filtrate concentration, low-temperature grinding are extremely Particle diameter is that 0.1-0.3mm i.e. obtains protective agent；

3) fermentation, homogenizing breaking cellular wall: to step 2) fermentation medium adds the active extra dry red wine yeast powder of its quality 0.2-0.4% Controlling temperature 30-34 DEG C, speed of agitator 50-100r/min, ferment at constant temperature 12-15h, then with the speed of 0.8-1.0 DEG C/min Rate is warming up to 40-42 DEG C, adds the active extra dry red wine yeast powder constant temperature static fermentation 3-5h of fermentation medium quality 0.1-0.3%, Finally be cooled to 28-32 DEG C with the speed of 0.6-0.8 DEG C/min, continue static fermentation 15-20h, fermentation liquid successively through 40 mesh, 80 mesh duplex strainers filter, filtrate through superhigh-voltage homogenizing machine in 2-4 DEG C, 140-160MPa homogenizing breaking cellular wall obtain homogenizing fluid；

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 1, it is characterised in that step 1) described yellow peach tank Head leftover bits and pieces is: any one or two kinds in yellow peach peels, inferior yellow peach sarcocarp mix with any mass ratio.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 1, it is characterised in that step 2) described microwave spoke Extract according to being extracted as batch (-type), i.e. microwave exposure 10s, be spaced 20s.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 1, it is characterised in that step 3) described activity do Rhodotorula glutinis powder is to prepare according to a conventional method with ocean rhodotorula CCTCC No:M 2014592 for the bacterium that sets out.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 4, it is characterised in that described activity extra dry red wine yeast powder The protective agent used in preparation process is step 2) prepare.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 1, it is characterised in that step 4) described vacuum from Heart condition is temperature 12-18 DEG C, vacuum-0.01--0.03MPa.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 1, it is characterised in that step 4) described biology break Emulsion is that glycolipid class, lipopeptid class, cell wall combine the one or more combination in class biological demulsifying agent.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 7, it is characterised in that step 4) described biology break The quality group of Emulsion becomes: glycolipid class: lipopeptid class: cell wall combines class=3-5:2-4:1-3.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 7 or 8, it is characterised in that described sugar lipidic biomass Demulsifier is one or both combinations in rhamnolipid, alkyl polyglucoside.

The method of a kind of fermenting and producing carotenoid the most as claimed in claim 9, it is characterised in that described sugar lipidic biomass breaks The quality group of Emulsion becomes: rhamnolipid: alkyl polyglucoside=4-6:1-2.