CN108004272A - A kind of method and lycopene product that lycopene is prepared using trispore Bruce mould - Google Patents

A kind of method and lycopene product that lycopene is prepared using trispore Bruce mould Download PDF

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CN108004272A
CN108004272A CN201711471926.9A CN201711471926A CN108004272A CN 108004272 A CN108004272 A CN 108004272A CN 201711471926 A CN201711471926 A CN 201711471926A CN 108004272 A CN108004272 A CN 108004272A
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coffee bean
lycopene
pericarp meat
bean pericarp
hydrolyzate
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CN108004272B (en
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李翔宇
陆姝欢
付杰
余超
汪志明
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Cabio Biotech Wuhan Co Ltd
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Cabio Biotech Wuhan Co Ltd
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
    • C12P5/007—Preparation of hydrocarbons or halogenated hydrocarbons containing one or more isoprene units, i.e. terpenes

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Abstract

The invention discloses a kind of method and lycopene product that lycopene is prepared using trispore Bruce mould, it is related to technical field of biological fermentation.The method provided by the invention for preparing lycopene, it includes：During fermented and cultured, coffee bean pericarp meat hydrolyzate is added into the zymotic fluid for be inoculated with trispore Bruce mould.This method is not to contain the coffee bean pericarp meat hydrolyzate of the intense irritation material such as nicotine, imidazoles as the blocking agent of cyclase, but the nitrogen heterocyclic ring alkaloids substance contained using coffee bean pericarp meat hydrolyzate blocks the generation of cyclase, reduce the residual of product moderate stimulation material, and promote the generation of cyclase, effectively improve lycopene product yield and quality.

Description

A kind of method and lycopene that lycopene is prepared using trispore Bruce mould Product
Technical field
The present invention relates to technical field of biological fermentation, and trispore Bruce mould preparation kind is utilized in particular to one kind The method and lycopene product of Lycopene.
Background technology
Three spore cloth Laplaces are mould to be currently mainly used in bata-carotene, has mechanism study to be shown to be due to the three mould hairs of spore cloth Laplace The synthesis of cyclase has blocked the generation of lycopene during ferment, so that product is mainly bata-carotene, is reported with many researchs Road using the intense irritation such as nicotine, imidazoles material come the synthesis of cyclase during blocking fermentation, so as to effectively facilitate tomato The synthesis of red pigment, but after the intense irritation material such as nicotine, imidazoles is utilized by thalline, product residual quantity can not be controlled effectively, Seriously affect the quality of lycopene product.
The content of the invention
It is an object of the invention to provide a kind of method that lycopene is prepared using trispore Bruce mould, this method with Blocking agent of the coffee bean pericarp meat hydrolyzate of the intense irritation material such as nicotine, imidazoles as cyclase is not contained, reduces production The residual of product moderate stimulation material, improves the quality of lycopene product.
It is a kind of by the above-mentioned preparation-obtained lycopene product of method another object of the present invention is to provide,.
What the present invention was realized in：
A kind of method for preparing lycopene, it includes：It is past to be inoculated with trispore Bruce mould during fermented and cultured Zymotic fluid in add coffee bean pericarp meat hydrolyzate.
A kind of lycopene product, its as it is above-mentioned prepare the method for lycopene prepared by obtain.
The invention has the advantages that：
The method provided by the invention for preparing lycopene, lycopene, this method are prepared using trispore Bruce mould Not contain the coffee bean pericarp meat hydrolyzate of the intense irritation material such as nicotine, imidazoles as the blocking agent of cyclase, but The nitrogen heterocyclic ring alkaloids substance contained using coffee bean pericarp meat hydrolyzate blocks the generation of cyclase, reduces and is pierced in product Swash the residual of property material, and promote the generation of cyclase, effectively improve lycopene product yield and quality.
Embodiment
, below will be in the embodiment of the present invention to make the purpose, technical scheme and advantage of the embodiment of the present invention clearer Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, builds according to normal condition or manufacturer The condition of view carries out.Reagents or instruments used without specified manufacturer, is the conventional production that can be obtained by commercially available purchase Product.
Below to a kind of method and tomato red that lycopene is prepared using trispore Bruce mould of the embodiment of the present invention Cellulose product is specifically described.
On the one hand, an embodiment of the present invention provides a kind of method for preparing lycopene, it includes：In fermented and cultured process In, coffee bean pericarp meat hydrolyzate is added into the zymotic fluid for be inoculated with trispore Bruce mould.
The present inventor has found first under study for action, by the clipped processing of coffee bean pericarp meat, acidolysis processing and Coffee bean pericarp meat hydrolyzate is obtained after enzymolysis processing and contains alkaloids substance, on the one hand, it can be used as with three spore cloth Stop the blocking agent that cyclase produces in Laplace mold fermentation production lycopene, avoid containing intense stimulus such as nicotine, imidazoles Property material addition, improve product quality, on the other hand, it can effectively facilitate mevalonic acid metabolic pathway, beneficial to tomato red The synthesis of element, improves yield of lycopene.
Further, in some embodiments of the present invention, during the 15-25h that ferments, coffee is added into zymotic fluid Coffee beans pericarp meat hydrolyzate.
Yield of lycopene can be improved by adding coffee bean pericarp meat hydrolyzate in the suitable fermentation time stage.Fermenting During 15-25h, coffee bean pericarp meat hydrolyzate is added into zymotic fluid, lycopene can be promoted to synthesize, improve its production Amount.
Further, in some embodiments of the present invention, the addition of coffee bean pericarp meat hydrolyzate is zymotic fluid The 4-8% of volume.
It should be noted that coffee bean pericarp meat hydrolyzate is added in a manner of once adding during the 15-25h that ferments Enter into zymotic fluid.
Further, in some embodiments of the present invention, in 30h to during fermentation ends, with 0.8-1.5g L-1·h-1Flow velocity toward adding coffee bean pericarp meat hydrolyzate in zymotic fluid.
In 30h to during fermentation ends, with 0.8-1.5gL-1·h-1Flow velocity toward adding coffee bean fruit in zymotic fluid Skin and flesh hydrolyzate, can further promote mevalonic acid metabolic pathway, improve the yield of lycopene.
Further, in some embodiments of the present invention, coffee bean pericarp meat hydrolyzate is prepared via a method which Obtain：
The hydrolysis of coffee bean pericarp meat will be obtained after the clipped processing of coffee bean pericarp meat, acidolysis processing and enzymolysis processing Liquid.
Further, in some embodiments of the present invention, the shear rate of shear treatment is 4000-8000r/min, Shear time is 5-10min.
Further, in some embodiments of the present invention, acidolysis, which is handled, is：It is the ratio of 1-5% by mass percentage Example, sour such as hydrochloric acid is added into the coffee bean pericarp meat after clipped processing), and the water with sour equivalent is added, it is uniformly mixed Afterwards, room temperature acidolysis 4-8h.
Further, in some embodiments of the present invention, enzymolysis processing is：It is the ratio of 1-5% by mass percentage , enzyme is added in the coffee bean pericarp meat toward after handling acidolysis, is placed under the conditions of 55-65 DEG C, digests 4-8h；
Enzyme includes alkali protease and cellulase.
Wherein, the enzyme activity of alkali protease be 1,500,000 U/g, the enzyme activity of cellulase be 1,000,000 U/g.
Further, in some embodiments of the present invention, in enzyme, the mass ratio of alkali protease and cellulase For：(3-4):(6-7).
On the other hand, an embodiment of the present invention provides a kind of lycopene product, its preparation kind stated by any of the above-described Obtained prepared by the method for Lycopene.
Lycopene product provided in an embodiment of the present invention has the features such as lycopene content is high, and impurity is few, and quality is high.
The feature and performance of the present invention are described in further detail with reference to embodiments.
Embodiment 1
The method provided in this embodiment for preparing lycopene, it includes the following steps：
1 prepares coffee bean pericarp meat hydrolyzate
1.1 shear treatment：Coffee bean pericarp meat 1000g is taken, is placed in high-speed shearing machine, is cut under 8000r/min speed Cut 10min；Obtain coffee bean pericarp digested tankage particle.
1.2 acidolysis are handled：It is 2% ratio by mass percentage, 10mol/L is added into coffee bean pericarp digested tankage particle Hydrochloric acid, and the water with sour equivalent is added, after mixing, it is placed under room temperature, acidolysis 4h；Obtain coffee bean pericarp carnic acid solution Liquid.
1.3 enzymolysis processing：It is 2% ratio (enzyme by mass percentage by enzyme：Coffee bean pericarp digested tankage particle=2%) plus Enter to by coffee bean pericarp meat acid hydrolysis solution, be placed under the conditions of 60 DEG C, digest 4h；Up to coffee bean pericarp meat hydrolyzate.
Wherein, enzyme is made of alkali protease and cellulase, and the weight ratio of alkali protease and cellulase is 3:7.
2 fermented and cultureds
2.1 inclined-plane culture
Take the positive bacterium of trispore Bruce mould and trispore Bruce mould to bear the spore suspension of bacterium, be respectively coated on PDA inclined-planes On culture medium, cultivated 5-7 days in 25 DEG C of constant incubators；
Wherein, the positive bacterium of trispore Bruce mould is trispore Bruce mould BT7251 (+), deposit number CCTCC M2014378；It is trispore Bruce mould BT7603 (-), deposit number CCTCC that trispore Bruce mould, which bears bacterium, M2014379。
2.2 seed culture
Respectively from the PDA slant mediums of the positive and negative bacterial strain of trispore Bruce mould with inoculation shovel shovel take a shovel positive bacterium, Negative bacterium, then be inoculated into respectively in the 1000ml triangular flasks containing 150ml seed culture mediums, under the conditions of 25 DEG C, 180 revs/min 48h is cultivated, the positive bacterium seed liquor of trispore Bruce mould is obtained and trispore Bruce mould bears bacterium seed liquor.
2.3 fermented and cultured
By the positive bacterium seed liquor of the trispore Bruce mould obtained in step 2.2 and trispore Bruce mould bear bacterium seed liquor by It is 1 according to positive and negative bacterium thalline mass ratio:5 are uniformly mixed, and are accessed with 10% (volume ratio) inoculum concentration in 50L fermentation tanks, fermented and cultured Process controls：25 DEG C of cultivation temperature, 300 revs/min of mixing speed, throughput 3vvm (L/L.min), tank pressure 0.1MPa, incubation time 120h, control in zymotic fluid concentration of glucose in 10-20g/ in fermentation process by stream plus glucose L。
Wherein, in fermentation 20h, coffee bean pericarp meat hydrolyzate is disposably added into zymotic fluid, addition is fermentation The 5% of liquid product；
During 30-120h, with 1gL-1·h-1Flow velocity toward adding coffee bean pericarp meat hydrolyzate in zymotic fluid.
3 detections
After fermentation, precise 0.02g dry myceliums, are extracted with tetrahydrofuran, with high performance liquid chromatography detection kind Lycopene yield, the results are shown in Table 2.
Wherein, high-efficiency liquid chromatography method for detecting is with reference to as follows：
(1) chromatographic condition：
Chromatographic column：Suplex PKB-100(Supelco)；250 × 4.6mm, 5 μm；
Wavelength：472nm；
Flow velocity：0.5ml/min；
Sampling volume：10μL；
Column temperature：30℃；
(2) mobile phase preparation and condition：
A phases：Methanol
B phases：50mg BHT are weighed into 1L volumetric flasks, add 20ml isopropanols to dissolve.Add 0.2ml N, N- diisopropyls Ethamine, 0.2% ammonium acetate solutions of 25ml, 455ml acetonitriles, 450ml methanol, solution recover to room temperature, and are extremely carved with methanol dilution Degree.
Eluent gradient table
4 use existing common process, such as using the lycopene in solvent extraction thalline, further precipitation, purifying, After crystallization, you can the lycopene product of high-purity is made.
Wherein, in other examples, step 3-4 is optional step, and the producer can select to carry out according to actual conditions.
Embodiment 2
The method provided in this embodiment for preparing lycopene, it includes the following steps：
1 prepares coffee bean pericarp meat hydrolyzate
1.1 shear treatment：Coffee bean pericarp meat 800g is taken, is placed in high-speed shearing machine, in 4000r/min speed down cuts 10min；Obtain coffee bean pericarp digested tankage particle.
1.2 acidolysis are handled：It is 2% ratio by mass percentage, 10mol/L is added into coffee bean pericarp digested tankage particle Hydrochloric acid, and the water with sour equivalent is added, after mixing, it is placed under room temperature, acidolysis 4h；Obtain coffee bean pericarp carnic acid solution Liquid.
1.3 enzymolysis processing：It is 2% ratio (enzyme by mass percentage by enzyme：Coffee bean pericarp digested tankage particle=2%) plus Enter to by coffee bean pericarp meat acid hydrolysis solution, be placed under the conditions of 60 DEG C, digest 4h；Up to coffee bean pericarp meat hydrolyzate.
Wherein, enzyme is made of alkali protease and cellulase, and the weight ratio of alkali protease and cellulase is 4:6.
2 fermented and cultureds
With embodiment 1
Unlike, during the fermentation, in fermentation 24h, coffee bean pericarp meat water is disposably added into zymotic fluid Liquid is solved, addition is the 6% of fermentating liquid volume；
During 30-120h, with 0.9gL-1·h-1Flow velocity toward adding the hydrolysis of coffee bean pericarp meat in zymotic fluid Liquid.
Remaining is the same as embodiment 1.
Embodiment 3
The method provided in this embodiment for preparing lycopene, it includes the following steps：
1 prepares coffee bean pericarp meat hydrolyzate
1.1 shear treatment：Coffee bean pericarp meat 800g is taken, is placed in high-speed shearing machine, in 6000r/min speed down cuts 8min；Obtain coffee bean pericarp digested tankage particle.
1.2 acidolysis are handled：It is 5% ratio by mass percentage, 10mol/L is added into coffee bean pericarp digested tankage particle Hydrochloric acid, and the water with sour equivalent is added, after mixing, it is placed under room temperature, acidolysis 7h；Obtain coffee bean pericarp carnic acid solution Liquid.
1.3 enzymolysis processing：It is 5% ratio (enzyme by mass percentage by enzyme：Coffee bean pericarp digested tankage particle=5%) plus Enter to by coffee bean pericarp meat acid hydrolysis solution, be placed under the conditions of 65 DEG C, digest 8h；Up to coffee bean pericarp meat hydrolyzate.
Wherein, enzyme is made of alkali protease and cellulase, and the weight ratio of alkali protease and cellulase is 5:5.
2 fermented and cultureds
With embodiment 1
Unlike, during the fermentation, in fermentation 15h, coffee bean pericarp meat water is disposably added into zymotic fluid Liquid is solved, addition is the 8% of fermentating liquid volume；
During 30-120h, with 1.2gL-1·h-1Flow velocity toward adding the hydrolysis of coffee bean pericarp meat in zymotic fluid Liquid.
Remaining is the same as embodiment 1.
Embodiment 4
The method provided in this embodiment for preparing lycopene, it includes the following steps：
1 prepares coffee bean pericarp meat hydrolyzate
1.1 shear treatment：Coffee bean pericarp meat 100g is taken, is placed in high-speed shearing machine, in 8000r/min speed down cuts 5min；Obtain coffee bean pericarp digested tankage particle.
1.2 acidolysis are handled：It is 1% ratio by mass percentage, 10mol/L is added into coffee bean pericarp digested tankage particle Hydrochloric acid, and the water with sour equivalent is added, after mixing, it is placed under room temperature, acidolysis 8h；Obtain coffee bean pericarp carnic acid solution Liquid.
1.3 enzymolysis processing：It is 3% ratio (enzyme by mass percentage by enzyme：Coffee bean pericarp digested tankage particle=3%) plus Enter to by coffee bean pericarp meat acid hydrolysis solution, be placed under the conditions of 65 DEG C, digest 8h；Up to coffee bean pericarp meat hydrolyzate.
Wherein, enzyme is made of alkali protease and cellulase, and the weight ratio of alkali protease and cellulase is 3.5: 6.5。
2 fermented and cultureds
With embodiment 1
Unlike, during the fermentation, in fermentation 18h, coffee bean pericarp meat water is disposably added into zymotic fluid Liquid is solved, addition is the 5% of fermentating liquid volume；
During 30-120h, with 1.2gL-1·h-1Flow velocity toward adding the hydrolysis of coffee bean pericarp meat in zymotic fluid Liquid.
Remaining is the same as embodiment 1.
Embodiment 5
The method provided in this embodiment for preparing lycopene, it includes the following steps：
1 prepares coffee bean pericarp meat hydrolyzate
With embodiment 1.
2 fermented and cultureds
With embodiment 1,
Unlike embodiment, during the fermentation, in fermentation 25h, coffee bean is disposably added into zymotic fluid Pericarp meat hydrolyzate, addition are the 7% of fermentating liquid volume；
During 30-120h, with 1.5gL-1·h-1Flow velocity toward adding the hydrolysis of coffee bean pericarp meat in zymotic fluid Liquid.
Remaining is the same as embodiment 1.
Comparative example 1
The method for preparing lycopene of this comparative example, including it is as follows：
Fermented and cultured is substantially the same manner as Example 1, unlike：
Wherein, in fermentation 20h, nicotine is disposably added into zymotic fluid, addition is the 0.15% of fermentating liquid volume.
Following detection step is the same as embodiment 1.
Comparative example 2
The method for preparing lycopene of this comparative example, including it is as follows：
Fermented and cultured is substantially the same manner as Example 1, unlike：
Wherein, in fermentation 25h, imidazoles is disposably added into zymotic fluid, addition is the 0.2% of fermentating liquid volume.
Following detection step is the same as embodiment 1.
Comparative example 3
The method for preparing lycopene of this comparative example, it is substantially the same manner as Example 1, the difference is that in fermented and cultured process In, in fermentation 30h, coffee bean pericarp meat hydrolyzate is disposably added into zymotic fluid, addition is fermentating liquid volume 10%；
During 30-120h, with 2gL-1·h-1Flow velocity toward adding coffee bean pericarp meat hydrolyzate in zymotic fluid.
Remaining is the same as embodiment 1.
Comparative example 4
The method for preparing lycopene of this comparative example, it is substantially the same manner as Example 1, the difference is that in fermented and cultured process In, in fermentation 10h, coffee bean pericarp meat hydrolyzate is disposably added into zymotic fluid, addition is fermentating liquid volume 2%；
During 30-120h, with 0.4gL-1·h-1Flow velocity toward adding the hydrolysis of coffee bean pericarp meat in zymotic fluid Liquid.
Remaining is the same as embodiment 1.
Lycopene content in the lycopene biological products of 1 embodiment 1-5 of table
In table：The lycopene content of dry mycelium refers to every 100g dry myceliums amount containing lycopene (g).
As can be seen from Table 1, the lycopene that the method for preparing lycopene that embodiment 1-5 is provided obtains contains Amount applies the product that the method for preparing lycopene that a 1-5 is provided obtains and is substantially free of nicotine apparently higher than comparative example 1-3 And imidazoles, greatly increase lycopene product quality.
To sum up, it is residual in product after the intense irritation material such as the nicotine as cyclase blocking agent, imidazoles is utilized by thalline Allowance can not be controlled effectively, seriously affect lycopene product quality, and the present invention utilizes coffee bean processing byproduct coffee bean fruit Skin and flesh carries out acidolysis and enzymolysis after high speed shear, obtains coffee bean pericarp meat hydrolyzate, wherein, except containing abundant ammonia Base nitrogen can promote thalli growth to utilize, and more have a certain amount of nitrogen heterocyclic ring and alkaloids substance, these materials were fermenting The synthesis of cyclase can be effectively blocked in journey, so as to improve yield of lycopene.
In short, the method provided by the invention for preparing lycopene uses coffee bean pericarp meat hydrolyzate as raw material preparation kind Lycopene has cost low, and raw material sources security quality is high, and accessory substance, which recycles, to economize on resources, yield of lycopene and quality The features such as high.
The foregoing is only a preferred embodiment of the present invention, is not intended to limit the invention, for the skill of this area For art personnel, the invention may be variously modified and varied.Within the spirit and principles of the invention, that is made any repaiies Change, equivalent substitution, improvement etc., should all be included in the protection scope of the present invention.

Claims (10)

  1. A kind of 1. method for preparing lycopene, it is characterised in that it includes：It is past to be inoculated with three spores during fermented and cultured Coffee bean pericarp meat hydrolyzate is added in the zymotic fluid of cloth Laplace mould.
  2. 2. the method according to claim 1 for preparing lycopene, it is characterised in that
    During the 15-25h that ferments, the coffee bean pericarp meat hydrolyzate is added into the zymotic fluid.
  3. 3. the method according to claim 2 for preparing lycopene, it is characterised in that the coffee bean pericarp meat hydrolyzate Addition be the fermentating liquid volume 4-8%.
  4. 4. the method for preparing lycopene according to Claims 2 or 3, it is characterised in that
    In 30h to during fermentation ends, with 0.8-1.5gL-1·h-1Flow velocity toward adding coffee bean fruit in the zymotic fluid Skin and flesh hydrolyzate.
  5. 5. the method according to any one of claim 1-3 for preparing lycopene, it is characterised in that the coffee bean fruit Skin and flesh hydrolyzate is prepared via a method which to obtain：
    The coffee bean pericarp meat hydrolysis will be obtained after the clipped processing of coffee bean pericarp meat, acidolysis processing and enzymolysis processing Liquid.
  6. 6. the method according to claim 5 for preparing lycopene, it is characterised in that the shear rate of the shear treatment For 4000-8000r/min, shear time 5-10min.
  7. 7. the method according to claim 5 for preparing lycopene, it is characterised in that the acidolysis, which is handled, is：By quality Percentage is the ratio of 1-5%, and acid is added into the coffee bean pericarp meat after clipped processing, and adds the water with sour equivalent, After mixing, room temperature acidolysis 4-8h.
  8. 8. the method according to claim 5 for preparing lycopene, it is characterised in that the enzymolysis processing is：By quality Percentage is the ratio of 1-5%, adds enzyme in the coffee bean pericarp meat toward after handling acidolysis, is placed under the conditions of 55-65 DEG C, enzyme Solve 4-8h；
    The enzyme includes alkali protease and cellulase.
  9. 9. the method according to claim 8 for preparing lycopene, it is characterised in that in the enzyme, alkali protease and The mass ratio of cellulase is：(3-4):(6-7).
  10. 10. a kind of lycopene product, it is characterised in that it any one of claim 1-9 as preparing lycopene Method prepared by obtain.
CN201711471926.9A 2017-12-28 2017-12-28 Method for preparing lycopene by utilizing Blakeslea trispora and lycopene product Active CN108004272B (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110283854A (en) * 2019-08-08 2019-09-27 内蒙古金达威药业有限公司 A kind of fermentation medium and its application and the method for preparing lycopene using Blakeslea trispora fermentation

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