CN110066839A - Culture medium for xanthan gum fermentation - Google Patents
Culture medium for xanthan gum fermentation Download PDFInfo
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- CN110066839A CN110066839A CN201910235431.9A CN201910235431A CN110066839A CN 110066839 A CN110066839 A CN 110066839A CN 201910235431 A CN201910235431 A CN 201910235431A CN 110066839 A CN110066839 A CN 110066839A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
- C12P19/06—Xanthan, i.e. Xanthomonas-type heteropolysaccharides
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Abstract
The invention belongs to fermentation technical field, the culture medium for xanthan gum fermentation is disclosed comprising following component: glucose, cornstarch, mycoprotein enzymolysis liquid, oleic acid, calcium carbonate, epsom salt, dipotassium hydrogen phosphate, fulvic acid, VB1.Culture medium cost of the present invention is cheap, and compared with prior art, fermentation produces the efficiency of xanthan gum and quality is promoted.
Description
Technical field
The invention belongs to fermentation technical fields, and in particular to the culture medium for xanthan gum fermentation.
Background technique
Xanthan gum (Xanthan gum), also known as Xanthan Gum or xanthan gum, be by cabbage black rot Xanthomonas campestris (
Xanthomonas campestris) or other Xanthomonas bacterial strain using carbohydrate as the fermented production of primary raw material
The extracellular heteroglycan of raw polymeric acidic, biosynthesis mechanism are " non-template synthesis mechanism ".Xanthan molecules are by " pentasaccharides
Repetitive unit " topology convergence body, xanthan molecules form bifilar deuterostrophies stereochemical structure, bifilar helix by hydrogen bond to each other
It is combined between structure by faint non-covalent bond, is arranged in the three-level spiral paradigmatic structure of neat " super bonding ribbon shape "；In pole
In the aqueous solution of dilute (< 1 %), typical level Four chrysanthemum shape aggregated structure is presented in xanthan gum when not heating.
Xanthan gum good water solubility sufficiently forms high viscosity solution after hydration, be the current thickening of collection in the world, suspend, emulsification,
It is stable at one, the biogum that best performance is got over；It can be used as emulsifier, stabilizer, gelling thickener, size, film forming agent
Deng；It is widely used in the fields such as food, medicine, chemical industry, petroleum.
Currently, the production method of xanthan gum mainly has fermentation method, protein Hydrolyze method and three kinds of chemical synthesis, wherein micro-
Biological fermentation process has become the main stream approach of production xanthan gum, how to optimize to fermentation medium and condition of culture,
It is our technical issues that need to address to improve yield and the quality of xanthan gum.
It is found by researches that the selection of carbon source and C/N compare fermenting speed, fermentation period has important influence, from logarithm
Growth period can promote the synthesis of xanthan gum using higher C/N ratio.The extension in same a batch xanthan gum fermentation period, third
Ketone acid content constantly increases.Therefore, C/N ratio is acted on the synthesis for passing to pyruvic acid by controlling the speed of growth.Application
Ren Fufeng group has also carried out a large amount of research to fermentation medium and culture parameters step, patented technology " CN103074408A,
A kind of production method of instant xanthan gum " discloses a kind of method for preparing xanthan gum using Xanthomonas campestris fermentation, wherein seed
Tank culture medium: glucose 2.5%, beef extract 3%, K2HPO43H2O0.2%, Na2HPO40.2%, biotin 0.5%,
MgSO47H2O0.2%, urea 0.025%, 115 DEG C of sterilizing 15min；Fermenter Medium Component: glucose 10%, corn
Starch 0.5%, beef extract 3%, MgSO47H2O 0.2%, K2HPO43H2O 0.2%, FeSO4 0.0001%, VB1
0.00001%, pH7.0~7.2.In above-mentioned incubation, mainly using beef extract as nitrogen source, higher cost also has other
It selects yeast extract etc. to be used as nitrogen source in research, equally exists defect at high cost.A kind of patented technology " work for preparing xanthan gum
Skill " discloses the preparation method of fermentation medium comprising following steps: taking glucose 9%, soybean egg according to weight percent
White 0.8%, corn pulp 2.5%, epsom salt 0.3%, dipotassium hydrogen phosphate 0.3%, potassium dihydrogen phosphate 0.1%, seven water sulfuric acid are sub-
Iron 0.001%, VB10.00001%, remaining is water, is stirred evenly, and pH6.5 is adjusted；The fermentation medium reduces costs, but
Be fermentation produce xanthan gum yield and quality still have it is to be hoisted.
Summary of the invention
It lacks present invention aim to address xanthan gum fermentation culture medium cost in the prior art is higher and yield is lower
It falls into, provides the culture medium for xanthan gum fermentation.
The present invention is achieved by the following technical solution:
Culture medium for xanthan gum fermentation comprising carbon source, nitrogen source, calcium carbonate.
Further,
The culture medium includes carbon source, nitrogen source, oleic acid, calcium carbonate.
Further,
The culture medium includes carbon source, nitrogen source, calcium carbonate, oleic acid, fulvic acid.
Further,
The culture medium includes following component: glucose, cornstarch, mycoprotein enzymolysis liquid, oleic acid, calcium carbonate, seven water sulphur
Sour magnesium, dipotassium hydrogen phosphate, fulvic acid, VB1。
Further,
The culture medium is grouped as by following group: glucose 40g/L, cornstarch 60g/L, mycoprotein enzymolysis liquid 50g/L, oil
Sour 10g/L, calcium carbonate 3g/L, epsom salt 1g/L, dipotassium hydrogen phosphate 1g/L, fulvic acid 20mg/L, VB120mg/L, pH
7.0-7.2。
Preferably,
The thallus enzymolysis liquid the preparation method comprises the following steps: collect the thallus in xanthan gum fermentation broth, it is dry to be less than to moisture content
The dry mycelium of 5wt%, being diluted with water to dry mycelium concentration is 40g/L, is placed in high-speed shearing machine and is cut with the speed of 10000rpm
120s is cut, bacteria suspension is obtained, the concentration that same volume is added into bacteria suspension is the hydrochloric acid solution of 1mol/L, is mixed, at 95 DEG C
Lower processing 1h adds trypsase later and is hydrolyzed, and then ceramic membrane filter, collects filtrate, then enzyme deactivation is condensed into dry
The paste that matter content is 40% is to get thallus enzymolysis liquid.
Preferably,
The hydrolysising condition of the trypsase are as follows: pH 8, temperature be 37 DEG C, hydrolysis time 6h.
Preferably,
The enzyme activity of the trypsase is 4000U/g.
Compared with prior art, the beneficial effect that the present invention obtains mainly includes but is not limited to the following aspects:
Carbon source a part is used to constitute cell component, and a part maintains normal metabolism, and another part is produced for purpose
The synthesis of object.The present invention uses glucose and cornstarch mixed carbon source, saves cost of material, and bacterial strain preferentially uses glucose,
With the increase of bacterial strain concentration, the enzymes such as the amylase of secretion increase, and can digest cornstarch as carbon source；This method is with Huang
The discarded mycoprotein of virgin rubber fermentation is raw material, and fermentation medium, raw material is made as organic nitrogen source after trypsin hydrolysis
Thallus is left after fermentation, low in cost, compared with being used as feed, albumen potency is higher, and benefit is more preferable, can directly reduce
Product benefit improves in industrial cost.The nitrogen sources such as yeast extract are substituted by addition thallus enzymolysis liquid, can greatly save and be fermented into
This.
Addition glutamic acid can increase the yield of xanthan gum in the medium, and mycoprotein enzymolysis liquid of the present invention contains a large amount of paddy
Propylhomoserin (accounts for 10% of total amino acid or more), can increase the yield of xanthan gum.Calcium and magnesium inorganic ions also can to thalli growth and
Product formation has an impact, and for magnesium elements to the irritating effect of thalli growth, calcium carbonate is the weight for influencing yield of xanthan gum and quality
The factor is wanted, the synthesis of exoprotein can be reduced under the conditions of suitable, improves the yield of xanthan gum, in addition, calcium carbonate also has
There is buffered fermentation liquid pH.
Containing groups such as a large amount of phenolic hydroxyl groups, carbonyls in fulvic acid, electrolysis degree is higher, can promote xanthan gum synthesis process
In to O2Utilization, further increase gum yield and xanthan gum quality.Oleic acid can reduce gas-liquid and pass oxygen resistance, improve oxygen and pass
Matter rate enhances system oxygen delivery capacity, improves xanthan gum yield, and without being additionally provided energy.
Detailed description of the invention
Fig. 1: influence of the oleic acid additive amount to yield of xanthan gum in culture medium；
Fig. 2: influence of the fulvic acid additive amount to yield of xanthan gum in culture medium.
Specific embodiment
Those skilled in the art can use for reference present disclosure, be suitably modified realization of process parameters.In particular, it should be pointed out that
All similar substitutions and modifications are apparent to those skilled in the art, they are considered as being included in this hair
It is bright.Product and method of the invention is described by preferred embodiment, and related personnel can obviously not depart from this hair
Product as described herein and method are modified in bright content, spirit and scope or appropriate changes and combinations, to realize and answer
Use the technology of the present invention.For a further understanding of the present invention, the following describes the present invention in detail with reference to examples.
Embodiment 1
Culture medium for xanthan gum fermentation comprising following component:
Glucose 40g/L, cornstarch 60g/L, mycoprotein enzymolysis liquid 50g/L, oleic acid 10g/L, calcium carbonate 3g/L, seven water sulphur
Sour magnesium 1g/L, dipotassium hydrogen phosphate 1g/L, fulvic acid 20mg/L, VB120mg/L, pH 7.0-7.2.
The culture medium the preparation method comprises the following steps:
Take each raw material, be successively added in water, stir evenly, adjust pH to get；
The thallus enzymolysis liquid the preparation method comprises the following steps: collect the thallus in xanthan gum fermentation broth, it is dry to be less than to moisture content
The dry mycelium of 5wt%, being diluted with water to dry mycelium concentration is 40g/L, is placed in high-speed shearing machine and is cut with the speed of 10000rpm
120s is cut, bacteria suspension is obtained, the concentration that same volume is added into bacteria suspension is the hydrochloric acid solution of 1mol/L, is mixed, at 95 DEG C
Lower processing 1h adds trypsase later and is hydrolyzed, then ceramic membrane filter, collection filtrate, 90 DEG C of enzyme deactivation 10min, then
Be condensed into dry matter content be 40%(weight ratio) paste to get；The molecular cut off of ceramic membrane is 10000Da；It filters off
Except the macromolecular substances for being difficult to be utilized by bacterial strain, including cell-wall components, high molecular weight protein etc..
The hydrolysising condition of the trypsase are as follows: pH 8, temperature be 37 DEG C, hydrolysis time 6h；The tryptose
The enzyme activity of enzyme is 4000U/g, additive amount are as follows: enzyme-to-substrate dry mass ratio is 1:30.
Embodiment 2
The technique for preparing xanthan gum using above-mentioned culture medium fermentation comprising following steps:
According to 17915 seed liquor (1 × 10 of Xanthomonas campestris ATCC8CFU/mL) culture is accessed according to the inoculum concentration of 8% (volume ratio)
Fermented and cultured is carried out in base, 30 DEG C of temperature, fermented incubation time 72 hours, obtains fermentation liquid；During fermented and cultured, pass through tune
Saving speed of agitator and ventilatory capacity holding dissolved oxygen level is 20%, is being not less than residual sugar control by auto-feeding glucose solution
2%.
Comparative example 1
Culture medium for xanthan gum fermentation comprising following component:
Glucose 100g/L, yeast extract 20g/L, oleic acid 10g/L, calcium carbonate 3g/L, epsom salt 1g/L, dipotassium hydrogen phosphate
1g/L, fulvic acid 20mg/L, VB120mg/L, pH 7.0-7.2.
Comparative example 2
Culture medium for xanthan gum fermentation comprising following component:
Glucose 40g/L, cornstarch 60g/L, mycoprotein enzymolysis liquid 50g/L, calcium carbonate 3g/L, epsom salt 1g/L,
Dipotassium hydrogen phosphate 1g/L, fulvic acid 20mg/L, VB120mg/L, pH 7.0-7.2.
Comparative example 3
Culture medium for xanthan gum fermentation comprising following component:
Glucose 40g/L, cornstarch 60g/L, mycoprotein enzymolysis liquid 50g/L, oleic acid 10g/L, calcium carbonate 3g/L, seven water sulphur
Sour magnesium 1g/L, dipotassium hydrogen phosphate 1g/L, VB120mg/L, pH 7.0-7.2.
Comparative example 4
Culture medium for xanthan gum fermentation comprising following component:
Glucose 40g/L, cornstarch 60g/L, mycoprotein enzymolysis liquid 50g/L, calcium carbonate 3g/L, epsom salt 1g/L,
Dipotassium hydrogen phosphate 1g/L, VB120mg/L, pH 7.0-7.2.
Embodiment 3
One, influence of the culture medium of embodiment 1 and comparative example 1-4 to xanthan gum.
Analyze influence of each culture medium to xanthan gum fermentation, zymotechnique reference implementation example 2.Specific data target is shown in Table 1:
Table 1
| Index | Embodiment 1 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 |
| Yield of xanthan gum g/L | 31.3 | 30.5 | 26.4 | 27.2 | 25.5 |
| Viscosity mPa/s | 6647 | 6598 | 5854 | 6130 | 5812 |
| Average molecular weight g/mol | 1.43×107 | 1.46×107 | 1.31×107 | 1.23×107 | 1.15×107 |
| Pyruvic acid functional group content % | 5.13 | 5.09 | 4.76 | 4.87 | 4.65 |
Conclusion: compared with comparative example 1, the present invention replaces common yeast extract using mycoprotein enzymolysis liquid, while using beautiful
Rice starch substitutes part glucose, the influence not big to the quality of fermentation production glue, and gum yield of the invention is compared with comparative example 1
Also certain raising；Compared with comparative example 2-4, the present invention is superior to comparative example 2-4 in terms of yield of xanthan gum and quality, can
See, the quality that oleic acid and fulvic acid produce glue and xanthan gum to fermentation has facilitation, and the two collaboration uses, and effect is more preferable.
Two, the influence of oleic acid and fulvic acid additive amount to yield of xanthan gum in culture medium.
The additive amount of oleic acid is set as 0,2.5,5,10,20,40(g/L), zymotechnique is referring to embodiment 2；Such as Fig. 1 institute
Show, with the increase of oleic acid additive amount, yield of xanthan gum is gradually increased, and when to be added to 10g/L, yield of xanthan gum reaches peak
Value, continues growing the additive amount of oleic acid, and the yield of xanthan gum has small size decline there is no increasing instead.
The additive amount of fulvic acid is set as 0,5,10,20,40,80(mg/L), zymotechnique is referring to embodiment 2；Such as Fig. 2 institute
Show, with the increase of fulvic acid additive amount, bacterial strain promotes the utilization rate of oxygen, and correspondingly, yield of xanthan gum increases therewith, wait increase
When being added to 20mg/L, yield of xanthan gum reaches peak value, continues growing the additive amount of oleic acid, there is no obviously change the yield of xanthan gum
Become, selects the additive amount of 20mg/L more appropriate.
Embodiment 3
Influence of the hydrolysis process of the present invention to amino acid content in mycoprotein enzymatic hydrolysis:
Control group is set,
Control group 1: not using high-speed shearing machine to handle, remaining is the same as embodiment 1；
Control group 2: use concentration that the mode of 6h is hydrolyzed for the hydrochloric acid of 5mol/L.
Sporoderm-broken rate, protein content and total free amino acid content are shown in Table 2:
Table 2
| Group | Embodiment 1 | Control group 1 | Control group 2 |
| Sporoderm-broken rate % | 96.5 | 82.9 | 69.1 |
| Total free amino acid content mg/g(dry mycelium) | 357.5 | 261.4 | 193.6 |
Conclusion: by table 2 as it can be seen that using high-speed shearing machine shear treatment 120s, thallus sporoderm-broken rate can be greatly improved, to mention
High hydrolysis efficiency；The present invention is pre-processed using dilute hydrochloric acid, and acid concentration used is smaller, then acid to amino acid extent of the destruction very
It is small, mild mode of action is used, then so as to retain the nutritive value of hydrolysate；1 thallus of embodiment of the present invention enzymatic hydrolysis
Total free amino acid content highest in liquid, it is easier to utilized by bacterial strain, and the content of total free amino acid Glutamic Acid compared with
Height can reach 12% or so, have preferable facilitation to xanthan gum yield, without additionally adding glutamic acid, drop in the medium
Low cost.
Listed above is only best specific embodiment of the invention.It is clear that the invention is not restricted to which above embodiments, may be used also
With there are many deformations.All changes that those skilled in the art directly can export or associate from present disclosure
Shape is considered as protection scope of the present invention.
Claims (8)
1. being used for the culture medium of xanthan gum fermentation comprising carbon source, nitrogen source, calcium carbonate.
2. culture medium according to claim 1, which is characterized in that the culture medium includes carbon source, nitrogen source, oleic acid, carbonic acid
Calcium.
3. culture medium according to claim 2, which is characterized in that the culture medium includes carbon source, nitrogen source, oleic acid, carbonic acid
Calcium, fulvic acid.
4. culture medium according to claim 3, which is characterized in that the culture medium includes following component: glucose, corn
Starch, mycoprotein enzymolysis liquid, oleic acid, calcium carbonate, epsom salt, dipotassium hydrogen phosphate, fulvic acid, VB1。
5. culture medium according to claim 4, which is characterized in that the culture medium is grouped as by following group: glucose
40g/L, cornstarch 60g/L, mycoprotein enzymolysis liquid 50g/L, oleic acid 10g/L, calcium carbonate 3g/L, epsom salt 1g/L,
Dipotassium hydrogen phosphate 1g/L, fulvic acid 20mg/L, VB120mg/L, pH 7.0-7.2.
6. according to right want 3-5 described in culture medium, which is characterized in that the thallus enzymolysis liquid the preparation method comprises the following steps: collecting yellow
Thallus in virgin rubber fermentation liquid, the dry dry mycelium for being less than 5wt% to moisture content, being diluted with water to dry mycelium concentration is 40g/
L is placed in high-speed shearing machine and shears 120s with the speed of 10000rpm, obtains bacteria suspension, same volume is added into bacteria suspension
Concentration be 1mol/L hydrochloric acid solution, mix, 1h is handled at 95 DEG C, later add trypsase be hydrolyzed, then make pottery
The filtering of porcelain film, collects filtrate, and enzyme deactivation is condensed into the paste that dry matter content is 40% finally to get thallus enzymolysis liquid.
7. according to right want 6 described in culture medium, which is characterized in that the hydrolysising condition of the trypsase are as follows: pH 8, temperature
Degree is 37 DEG C, hydrolysis time 6h.
8. culture medium according to claim 7, which is characterized in that the enzyme activity of the trypsase is 4000U/g.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110964762A (en) * | 2019-12-24 | 2020-04-07 | 内蒙古阜丰生物科技有限公司 | Fermentation process of low-starch-residue xanthan gum product |
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