CN102732049B - Method for preparing and extracting carotenoid from microbial thalli - Google Patents
Method for preparing and extracting carotenoid from microbial thalli Download PDFInfo
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- CN102732049B CN102732049B CN201210180281.4A CN201210180281A CN102732049B CN 102732049 B CN102732049 B CN 102732049B CN 201210180281 A CN201210180281 A CN 201210180281A CN 102732049 B CN102732049 B CN 102732049B
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Abstract
The invention which belongs to bioengineering and bioseparation engineering fields relates to a method for preparing and extracting carotenoid from microbial thalli. The method comprises the following steps: fermenting the microbial thalli, separating the resulting fermentation solution and the thalli to obtain wet thalli, guiding the wet thalli to an explosion tank through a material flow pipeline, and carrying out high-pressure explosion wall breaking; dehydrating the wet thalli with disrupted cells to obtain a filter cake formed by thallium fragments; adding 10-25L of an organic solvent to each 1kg of the filter cake, and carrying out stirring extraction at 30-55DEG C for 20-50min; filtering to obtain an organic solvent extraction containing carotenoid grease; and carrying out vacuum concentration on the extraction at 40-60DEG C, and recovering the organic solvent to obtain the carotenoid grease. The extraction yield one time reaches above 90%. The method has the advantages of omission of dehydration drying and thallium crushing operations in traditional technologies, short process flow, high extraction yield and short time each time, reduced organic solvent application amount, substantially reduced energy consumption and other costs, simple whole process, and controllable quality, and is suitable for industrialized production applications.
Description
Technical field
The invention belongs to biotechnology and bioseparation engineering field, be specifically related to a kind of method of preparing carotenoid of extracting from microbial cells.The method is particularly useful for a large amount of extraction from blakeslea trispora and phaffiafhodozyma and prepares carotenoid.
Background technology
β-carotene, gamma carotene, Lyeopene, astaxanthin are four kinds of main carotenoid in microorganism.β-carotene, Lyeopene, astaxanthin are the lipophilic compounds that marketable value is very high.In foodstuffs industry, be natural pigment, be antidotal antioxidant in cosmetic industry, is anticancer medicine in pharmaceutical industry.Food, makeup, pharmaceutical industries also increase day by day to the demand of these carotenoid, good market prospects.At present, Lyeopene mainly extracts from tomato, and β-carotene mainly extracts from salt algae, the under-supply maximum bottleneck becoming in production of raw material.Fermentation method handiness aspect production time, region selection, raw material supply is very high, is the Perfected process of producing carotenoid.
Blakeslea trispora is the major microorganisms of current suitability for industrialized production β-carotene and Lyeopene, and phaffiafhodozyma is the major microorganisms of fermentative production astaxanthin.Carotenoid is thin intracellular product.Therefore, the extraction of carotenoid comprise cell fragmentation, dehydrate, the step such as organic solvent extraction and concentration and recovery.As used vacuum-drying to make mycelium dehydration in patent application 200910135676.0 " method of preparing β-carotene from Blakeslea trispora fermentation broth ", grind dry mycelium, dichloromethane extraction.In patent application 200910158159.5 " by the natural beta-carotin of Blakeslea trispora fermentative production or the sample treatment of Lyeopene fermented liquid ", use ultrasonication, dehydrated alcohol dehydration, ethyl acetate to extract.These extracting method all need to dehydrate and thalline pulverizing process, and technical process is long, single extract yield is low and large with duration, organic solvent consumption, and energy consumption is high with other costs, and industrial applications is restricted.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of method of preparing carotenoid of extracting from microbial cells is provided.The method is saved dehydrating and thalline pulverizing process in traditional extraction technique, and technical process is short, and single extract yield is high and the used time is short, organic solvent consumption reduces, and energy consumption and other costs significantly reduce, and whole extraction preparation process is simple, quality controllable, be suitable for suitability for industrialized production application.
A kind of method of preparing carotenoid of extracting from microbial cells provided by the invention, comprises the following steps:
(1) after microbial cells fermentation ends, fermented liquid is separated with thalline, obtains wet thallus;
(2) wet thallus imports in cavity charge containers by logistics pipeline, adopts steam explosion method broken wall, makes wet thallus cytoclasis;
(3) by the wet thallus dehydration after cytoclasis, obtain the filter cake that bacterial chip forms;
(4) filter cake is joined in organic solvent, every kilogram of filter cake needs 10～25 liters of organic solvents, and 30～55 ℃ of stirring and leaching 20～50 minutes are filtered, and obtain the organic solvent extracting solution containing carotenoid grease;
(5) extracting solution vacuum concentration under 40～60 ℃, vacuum tightness 0.01～0.1MPa condition, reclaims organic solvent, obtains the grease containing carotenoid.
As the improvement of technique scheme, in step (1) the fermented liquid method separated with thalline be staticly settle, Plate Filtration or centrifugal.
As the improvement of technique scheme, steam explosion method described in step (2) is that thalline is steam heated to saturated vapor pressure 0.1～0.5MPa explosion with 0.1～0.8MPa.
As the further improvement of technique scheme, in step (3), the method for dehydration is vacuum filtration, Plate Filtration or centrifugal, obtains the filter cake that bacterial chip forms.
As the further improvement of technique scheme, organic solvent is acetone or alcohol acetone mixed solution in step (4), and in ethanol acetone mixed solution, ethanol volume percent content can be between 0 to 50%.
Described microbial cells is particularly useful for blakeslea trispora and phaffiafhodozyma.
A kind of methods of preparing carotenoid of extracting in a large number from microbial cells provided by the invention, compared with prior art, have following advantage:
Method of the present invention is saved dehydrating and thalline pulverizing process in traditional extraction technique, and technical process is short, and single extract yield is high and the used time is short, and organic solvent consumption reduces, and energy consumption and other costs significantly reduce.Whole extraction preparation process is simple, quality controllable, is suitable for suitability for industrialized production application.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
Embodiment 1
After blakeslea trispora fermentation ends, staticly settle, abandon supernatant liquor, obtain wet thallus.Wet thallus imports in cavity charge containers by logistics pipeline, with 0.3MPa, be steam heated to saturated vapor pressure 0.1MPa steam explosion, the centrifugal thalline filter cake that obtains, get 200g thalline filter cake, use 5L acetone, 50 ℃ of stirring and leaching 30 minutes, filtration obtains the extracting solution of clarification, vacuum concentration under 50 ℃ and vacuum tightness 0.01MPa condition, reclaims acetone, obtains the carotenoid grease containing β-carotene, gamma carotene, Lyeopene.The rate of recovery of β-carotene reaches 90%, and the rate of recovery of gamma carotene reaches 93%, and the rate of recovery of Lyeopene reaches 92%.
Embodiment 2
After blakeslea trispora fermentation ends, the centrifugal wet thallus that obtains.Wet thallus imports in cavity charge containers by logistics pipeline, with 0.1MPa, be steam heated to saturated vapor pressure 0.2MPa steam explosion, vacuum filtration obtains thalline filter cake, get 2Kg wet thallus filter cake, with 50L ethanol acetone mixed solvent (ethanol volume percent content is 30%), 35 ℃ of stirring and leaching 50 minutes, filtration obtains the extracting solution of clarification, vacuum concentration under 40 ℃ and vacuum tightness 0.05MPa condition, reclaim organic solvent, obtain the carotenoid grease containing β-carotene, gamma carotene, Lyeopene.The rate of recovery of β-carotene reaches 93%, and the rate of recovery of gamma carotene reaches 92%, and the rate of recovery of Lyeopene reaches 95%.
Embodiment 3
After blakeslea trispora fermentation ends, staticly settle, abandon supernatant liquor, obtain wet thallus.Wet thallus imports in cavity charge containers by logistics pipeline, with 0.6MPa, be steam heated to saturated vapor pressure 0.15MPa steam explosion, Plate Filtration obtains thalline filter cake, get 20Kg wet thallus filter cake, with 300L ethanol acetone mixed solvent (ethanol volume percent content is 50%), 40 ℃ of stirring and leaching 45 minutes, filtration obtains the extracting solution of clarification, vacuum concentration under 60 ℃ and vacuum tightness 0.08MPa condition, reclaim ethanol acetone, obtain the carotenoid grease containing β-carotene, gamma carotene, Lyeopene.The rate of recovery of β-carotene reaches 93%, and the rate of recovery of gamma carotene reaches 93%, and the rate of recovery of Lyeopene reaches 94%.
Embodiment 4
After blakeslea trispora fermentation ends, Plate Filtration obtains wet thallus.Wet thallus imports in cavity charge containers by logistics pipeline, with 0.8MPa, be steam heated to saturated vapor pressure 0.2MPa steam explosion, vacuum filtration obtains thalline filter cake, get 200Kg wet thallus, with 2000L ethanol acetone mixed solvent (ethanol volume percent content is 25%), 55 ℃ of stirring and leaching 30 minutes, filtration obtains the extracting solution of clarification, vacuum concentration under 50 ℃ and vacuum tightness 0.1MPa condition, reclaim ethanol acetone, obtain the carotenoid grease containing β-carotene, gamma carotene, Lyeopene.The rate of recovery of β-carotene reaches 90%, and the rate of recovery of gamma carotene reaches 91%, and the rate of recovery of Lyeopene reaches 92%.
Embodiment 5
After phaffiafhodozyma fermentation ends, the centrifugal wet thallus that obtains.Wet thallus imports in cavity charge containers by logistics pipeline again, with 0.8MPa, be steam heated to saturated vapor pressure 0.4MPa steam explosion, the centrifugal thalline filter cake that obtains, get 2kg wet thallus, with 20L ethanol acetone mixed solvent (ethanol volume percent content is 25%), 45 ℃ of stirring and leaching 20 minutes, the centrifugal extracting solution that obtains clarification, vacuum concentration under 50 ℃ and vacuum tightness 0.05MPa condition, reclaims ethanol acetone, obtains the carotenoid grease of astaxanthin-containing.The rate of recovery of astaxanthin reaches 93%.
Embodiment 6
After phaffiafhodozyma fermentation ends, staticly settle, abandon supernatant liquor, obtain wet thallus.Wet thallus imports in cavity charge containers by logistics pipeline again, with 0.8MPa, be steam heated to saturated vapor pressure 0.5MPa steam explosion, the centrifugal thalline filter cake that obtains, get 20Kg wet thallus, with 500L ethanol acetone mixed solvent (ethanol volume percent content is 15%), 50 ℃ of stirring and leaching 50 minutes, filtration obtains the extracting solution of clarification, vacuum concentration under 60 ℃ and vacuum tightness 0.06MPa condition, reclaims ethanol acetone, obtains the carotenoid grease of astaxanthin-containing.The rate of recovery of astaxanthin reaches 91%.
Embodiment 7
After the red fermentation using bacteria of fixed nitrogen finishes, the centrifugal wet thallus that obtains.Wet thallus imports in cavity charge containers by logistics pipeline again, with 0.6M Pa, be steam heated to saturated vapor pressure 0.25M Pa steam explosion, Plate Filtration obtains thalline filter cake, get 10Kg wet thallus, use 250L acetone, 50 ℃ of stirring and leaching 30 minutes, filtration obtains the extracting solution of clarification, vacuum concentration under 60 ℃ and vacuum tightness 0.06MPa condition, reclaims acetone, obtains the carotenoid grease containing Lyeopene.The rate of recovery of Lyeopene reaches 85%.
Those skilled in the art will readily understand; the foregoing is only preferred embodiment of the present invention; not in order to limit the present invention, all any modifications of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (1)
1. from microbial cells, extract a method of preparing carotenoid, it is characterized in that, the method comprises the following steps:
(1) after microbial cells fermentation ends, fermented liquid is separated with thalline, obtains wet thallus; Described microbial cells is the red bacterium of blakeslea trispora, phaffiafhodozyma or fixed nitrogen;
(2) wet thallus imports in cavity charge containers by logistics pipeline, adopts steam explosion method broken wall, makes wet thallus cytoclasis; Described steam explosion method is that wet thallus is steam heated to saturated vapor pressure 0.1～0.5MPa with 0.1～0.8MPa;
(3) by the wet thallus dehydration after cytoclasis, obtain the filter cake that bacterial chip forms; The method of dehydration is vacuum filtration, filter press or centrifugal;
(4) filter cake is joined in organic solvent, every kilogram of filter cake needs 10～25 liters of organic solvents, and 30～55 ℃ of stirring and leaching 20～50 minutes are filtered, and obtain the organic solvent extracting solution containing carotenoid grease; Organic solvent is acetone ethanol mixed solution, and in acetone ethanol mixed solution, ethanol volume percent content is between 0 to 50%;
(5) extracting solution vacuum concentration under 40～60 ℃, Absolute truth reciprocal of duty cycle 0.01～0.1MPa condition, reclaims organic solvent, obtains the grease containing carotenoid.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103012230A (en) * | 2013-01-06 | 2013-04-03 | 大连医诺生物有限公司 | Novel process for high-effective extraction of carotenoid in Blakeslea trispora |
| CN109206938B (en) * | 2016-07-08 | 2019-10-15 | 天津工业大学 | A kind of natural brown dyestuff |
| CN107164254B (en) * | 2016-09-13 | 2020-05-15 | 湖北广济药业股份有限公司 | Microorganisms and uses thereof |
| CN108467875B (en) * | 2018-04-28 | 2021-02-05 | 郑州轻工业学院 | Method for preparing dihydroactinidiolide by microbial fermentation of carotenoid |
| CN108707631A (en) * | 2018-05-02 | 2018-10-26 | 莲花健康产业集团股份有限公司 | A kind of glutamic acid fermentation discards the reuse method of thalline |
| CN111675923B (en) * | 2020-06-24 | 2021-05-25 | 河南大农联生物工程有限公司 | Method for extracting gardenia pigment through steam explosion and application method of extracted pigment |
| CN111995880B (en) * | 2020-08-20 | 2022-04-29 | 宜昌东阳光生化制药有限公司 | Method for extracting biologically fermented carotenoid |
| CN114133348A (en) * | 2021-11-09 | 2022-03-04 | 湖北广济药业股份有限公司 | Method for extracting high-purity all-trans beta-carotene |
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