CN101724618A - Method for culturing coriolus versicolor induced oxalate decarboxylase by using straw carbon source - Google Patents
Method for culturing coriolus versicolor induced oxalate decarboxylase by using straw carbon source Download PDFInfo
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- CN101724618A CN101724618A CN200910200932A CN200910200932A CN101724618A CN 101724618 A CN101724618 A CN 101724618A CN 200910200932 A CN200910200932 A CN 200910200932A CN 200910200932 A CN200910200932 A CN 200910200932A CN 101724618 A CN101724618 A CN 101724618A
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Abstract
The invention relates to a method for culturing coriolus versicolor induced oxalate decarboxylase by using a straw carbon source, which comprises the following steps of: (1) soaking crushed straws in acid solution, and hydrolyzing the crushed straws at the temperature of between 100 and 150 DEG C; (2) preparing a fermentation medium, and adjusting the pH value to between 4.0 and 5.0; (3) taking a mycelium, inoculating the mycelium into a liquid seed culture medium, when a white mycelium group is formed but is not emerged on the water surface, transferring the mycelium and the culture medium to a container containing sterile glass beads, vibrating the container to break the mixture, and inoculating the mycelium into the fermentation medium according to the inoculum size of 3 to 15 percent; and (4), after 4 to 6 days of culture, adding oxalic acid into the container, performing the culture for 0.5 to 6 days to obtain the mycelium, grinding and crushing the mycelium after the mycelium is chilled by liquid nitrogen, extracting mycelial fragments by using an acetate buffer, and centrifuging the mycelial fragments. The method is simple, is easy in operation and is low in cost, the preparation of the carbon source has strong controllability, and the sugar content is stable.
Description
Technical field
The invention belongs to the preparation field of oxalate decarboxylase, particularly relate to the method that a kind of straw carbon source is cultivated the Corilus versicolor Quel. inducing oxalate decarboxylase.
Background technology
Oxalate decarboxylase can the decarboxylation of catalysis oxalic acid form formic acid and carbonic acid gas, thus in enzyme process detection oxalic acid content, degraded trade effluent medium-height grass hydrochlorate, aspects such as low oxalic acid content food of preparation and the interior concentration of oxalic acid of reduction human body have broad application prospects.How Corilus versicolor Quel. reduces it and cultivates cost as the important bacterial classification of producing oxalate decarboxylase, the mass production mycelium, and to induce mycelium synthesis of oxalic acid decarboxylase be an important topic.Because carbon is the maximum element of organism intensive amount, therefore carbon source is a kind of of consumption maximum in the substratum, the ratio that accounts for the biological product total cost is higher, so can reduce the cultivation cost of microorganism significantly by utilizing low-cost carbon source to substitute the pure glucose that uses at present.
Stalk is the harvested residuums of farm crop, and as the byproduct of agriculture production, annual a large amount of stalk is mainly used to make a fire in the rural area, perhaps pulverizes the back and does feed usefulness to domestic animal.Though the stalk after burning can be sprinkling upon the field as fertilizer sources, the gas that produces in burning process can cause the increase of Greenhouse effect, and the flue dust that produces also can cause atmospheric pollution.So the carbon source with the hydrolyzed solution that obtains after the stalk hydrolysis is used as cultivating industrial microorganism not only can reduce the microorganism culturing cost, alleviate the pollution of burning, and improved stalk and used added value environment, have great economic benefit and environmental benefit.
Summary of the invention
Technical problem to be solved by this invention provides the method that a kind of straw carbon source is cultivated the Corilus versicolor Quel. inducing oxalate decarboxylase, and this method is simple, and cost is low, the efficient height; Substrate for induction produces oxalate decarboxylase not to be suppressed, and yield of enzyme is stable.
A kind of straw carbon source of the present invention is cultivated the method for Corilus versicolor Quel. inducing oxalate decarboxylase, comprising:
(1) be 1g with the straw of pulverizing according to solid-to-liquid ratio: 5-15ml soaks 10-20h with acid solution, hydrolysis reaction 20-60min under 100-150 ℃ of condition, behind the centrifugal removal residue, obtain the straw hydrolyzed solution, and measure reducing sugar content in the hydrolyzed solution (with glucose as a standard sugar meter) with the DNS method;
(2) press the reducing sugar amount of 7-30g/L, the straw hydrolyzed solution is mixed with fermention medium, regulating pH value be 4.0-5.0, removes by filter impurity, and adds nitrogenous source and inorganic salt, reexamines and regulate the pH value to 4.0-5.0, and it is afterwards standby to sterilize;
(3) getting 2-3 sheet diameter is that 9～11mm contains mycelial agar solid medium, inserts liquid seed culture medium, leaves standstill in 30 ℃ and cultivates 4-6 days; Treat that mycelium liquid medium within top forms the white hypha group but when not emerging as yet, mycelium and substratum are all transferred in the container that contains sterile glass beads, mycelium is shaken the broken suspension liquid that obtains to contain the mycelia fragment, inoculum size with 3-15% inserts above-mentioned fermention medium, cultivates 4-6 days under 20-30 ℃ and 100-250rpm speed conditions;
When (4) treating that mycelium pellet forms, adding final concentration is the generation of the oxalic acid inducing oxalate decarboxylase of 5-20mM, cultivates and obtains mycelium after 0.5-6 days; Mycelium grinds broken behind liquid nitrogen freezing, uses 0.2M, and the acetate buffer solution of pH 3.7 extracts the mycelia fragment, centrifugal removal residue, and the gained supernatant liquor is the oxalate decarboxylase crude enzyme liquid.
Acid in the described step (1) is sulfuric acid, hydrochloric acid or acetic acid, and mass percent concentration is 0.5%-8%;
Described step (2) is regulated the pH value with sodium hydroxide solution;
The component of the fermention medium in the described step (2) is: the reducing sugar 7～30g in the straw hydrolyzate (with glucose as a standard sugar meter), peptone 3.0g, KH
2PO
41.0g, Na
2HPO
412H
2O 0.2g, MgSO
47H
2O 0.5g and micro-1ml, water is settled to 1L;
The component of described trace element is: FeSO
47H
2O 10g, MnSO
4H
2O 1.0g, ZnSO
47H
2O 1.0g, CuSO
45H
2O 2.0g, CaCl
22H
2O 13g, water is settled to 1L;
Liquid seed culture medium in the described step (3), with the reducing sugar in the glucose replacement fermention medium, all the other component concentrations are identical with fermention medium.
The present invention obtains to cultivate the carbon source that Corilus versicolor Quel. is grown by the hydrolysis straw, and induces mycelium synthesis of oxalic acid decarboxylase with substrate oxalic acid.
Beneficial effect
(1) the present invention utilizes the carbon source of straw hydrolyzed solution as the fermentation culture Corilus versicolor Quel., and method is simple, easy handling;
(2) the straw output of the present invention's use is big, and cost is low, and is cheap as the cost of material of production carbon source; And the preparation controllability of carbon source is strong, and sugar degree is stable;
(3) with traditional culture medium carbon source---glucose is compared, the mycelial biomass of producing the Corilus versicolor Quel. acquisition with this low value carbon source is big, the transformation efficiency height.
(4) compare with traditional substratum, when cultivating thalline with this low value carbon source, substrate for induction produces oxalate decarboxylase not to be suppressed, and yield of enzyme is stable.
Description of drawings
The mycelium dry weight that Fig. 1 straw hydrolyzed solution substratum (initial pH 5.0) is produced is with the variation of fermentation time;
The hydrolyzed solution substratum
Dextrose culture-medium;
Utilize oxalic acid to induce the synthetic enzyme activity in Fig. 2 straw hydrolyzed solution substratum (initial pH 5.0);
The hydrolyzed solution substratum
Dextrose culture-medium;
Add the mycelium protein content that oxalic acid induces the back to extract in Fig. 3 straw hydrolyzed solution substratum (initial pH 5.0);
The hydrolyzed solution substratum
Dextrose culture-medium;
Utilize oxalic acid to induce the ratio vigor 1 of synthetic enzyme in Fig. 4 straw hydrolyzed solution substratum (initial pH 5.0);
The hydrolyzed solution substratum
Dextrose culture-medium;
The mycelium crushing residua dry weight of Fig. 5 straw hydrolyzed solution substratum (initial pH 5.0) after mesoxalic acid is induced;
The hydrolyzed solution substratum
Dextrose culture-medium;
Utilize oxalic acid to induce the ratio vigor 2 of synthetic enzyme in Fig. 6 straw hydrolyzed solution substratum (initial pH 5.0);
The hydrolyzed solution substratum
Dextrose culture-medium;
Add the reducing sugar content after oxalic acid is induced in Fig. 7 straw hydrolyzed solution substratum (initial pH 5.0);
The hydrolyzed solution substratum
Dextrose culture-medium;
Fig. 8 straw hydrolyzed solution substratum (initial pH 5.0) mesoxalic acid induces the pH of secondary fermentation liquid to change;
The hydrolyzed solution substratum
Dextrose culture-medium;
The mycelium dry weight that Fig. 9 straw hydrolyzed solution substratum (initial pH 3.5) is produced.
The hydrolyzed solution substratum
Dextrose culture-medium
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
The straw hydrolyzed solution is produced Corilus versicolor Quel. (initial pH value of medium 5.0)
Abundant ground straw is weighed, adding concentration with the solid-to-liquid ratio of 1g: 12ml is 1% sulphuric acid soln, the straw powder is after dilution heat of sulfuric acid soaks 10 hours, be positioned in the high-pressure steam sterilizing pan, under 121 ℃, reaction 30min, remove by filter the straw residue, it is standby to obtain the straw hydrolyzed solution, and the content (with glucose as a standard sugar) with reducing sugar in the DNS method mensuration straw hydrolyzed solution stores for future use under 4 ℃ of conditions;
Add the straw hydrolyzed solution that contains 1g reducing sugar (with glucose meter) in every 50mL substratum, add the 0.15g peptone again, 0.05g potassium primary phosphate, 0.025g MgSO
47H
2O, 0.01g Na
2HPO
412H
2O, and micro-50 μ L (trace element formula is: FeSO
47H
2O 10g/L, MnSO
4H
2O 1.0g/L, ZnSO
47H
2O1.0g/L, CuSO
45H
2O 2.0g/L, CaCl
22H
2O 13g/L).After the substratum preparation is finished, regulate pH value to 5.0 with sodium hydroxide, the sterilization back is standby;
Get an agar plate bacterial classification, lay the about 10mm of diameter with punch tool and contain mycelial solid medium thin slice, get the 2-3 sheet and add liquid seed culture medium (replacing reducing sugar with glucose), 30 ℃ left standstill 4-6 days.Treat that mycelium liquid medium within top forms white cap but incomplete as yet emersion liquid level all moves into the containers that granulated glass sphere is housed, shake fragmentation, obtain to contain the suspension liquid of mycelia fragment.The bacteria suspension of mixing is inserted in the fermention medium by 10% inoculum size, cultivated 1-5 days, take by weighing mycelium over dry weight.The mycelium production of finding straw hydrolyzed solution substratum is higher than the mycelial output of control medium (glucose), and still is in ascendant trend, and 4～6 days is the growth animated period of mycelia.
The straw hydrolyzed solution is produced Corilus versicolor Quel. (initial pH value of medium 5.0)
Abundant ground straw is weighed, adding concentration with the solid-to-liquid ratio of 1g: 15ml is 3% sulphuric acid soln, the straw powder is after dilution heat of sulfuric acid soaks 10 hours, be positioned in the steaming and decocting under high pressure pot, under 100 ℃, reaction 60min, remove by filter the straw residue, it is standby to obtain the straw hydrolyzed solution, and the content (with glucose as a standard sugar) with reducing sugar in the DNS method mensuration straw hydrolyzed solution stores for future use under 4 ℃ of conditions;
Add the straw hydrolyzed solution that contains 1.5g reducing sugar (with glucose meter) in every 50mL substratum, add the 0.15g peptone again, 0.05g potassium primary phosphate, 0.025g MgSO
47H
2O, 0.01g Na
2HPO
412H
2O, and micro-50 μ L (trace element formula is: FeSO
47H
2O 10g/L, MnSO
4H
2O 1.0g/L, ZnSO
47H
2O1.0g/L, CuSO
45H
2O 2.0g/L, CaCl
22H
2O 13g/L).After the substratum preparation is finished, regulate pH value to 5.0 with sodium hydroxide, the sterilization back is standby;
Get an agar plate bacterial classification, lay the about 10mm of diameter with punch tool and contain mycelial solid medium thin slice, get the 2-3 sheet and add liquid seed culture medium, 30 ℃ left standstill 4-6 days.Treat that mycelium liquid medium within top forms white cap but incomplete as yet emersion liquid level all moves into the containers that granulated glass sphere is housed, shake fragmentation, obtain to contain the suspension liquid of mycelia fragment.The bacteria suspension of mixing is inserted in the fermention medium by 15% inoculum size, cultivated 1-5 days, take by weighing mycelium over dry weight.The variation tendency of finding mycelium production is similar to Example 1, is higher than control medium (glucose) mycelium production equally, and is in ascendant trend, and 4～6 days is the growth animated period of mycelia.
The straw hydrolyzed solution is produced Corilus versicolor Quel. (initial pH value of medium 5.0)
Abundant ground straw is weighed, and adding concentration with the solid-to-liquid ratio of 1g: 5ml is 8% sulphuric acid soln, and the straw powder is after dilution heat of sulfuric acid soaks 10 hours, be positioned in the steaming and decocting under high pressure pot, under 150 ℃, reaction 20min, remove by filter the straw residue, it is standby to obtain the straw hydrolyzed solution.Content (with glucose as a standard sugar) with reducing sugar in the DNS method mensuration straw hydrolyzed solution stores for future use under 4 ℃ of conditions;
Add the straw hydrolyzed solution that contains 0.35g reducing sugar (with glucose meter) in every 50mL substratum, add the 0.15g peptone again, 0.05g potassium primary phosphate, 0.025g MgSO
47H
2O, 0.01g Na
2HPO
412H
2O, and micro-50 μ L (trace element formula is: FeSO
47H
2O 10g/L, MnSO
4H
2O 1.0g/L, ZnSO
47H
2O 1.0g/L, CuSO
45H
2O 2.0g/L, CaCl
22H
2O 13g/L).After the substratum preparation is finished, regulate pH value to 5.0 with sodium hydroxide, the sterilization back is standby;
Get an agar plate bacterial classification, lay the about 10mm of diameter with punch tool and contain mycelial solid medium thin slice, get the 2-3 sheet and add liquid seed culture medium, 30 ℃ left standstill 4-6 days.Treat that mycelium liquid medium within top forms white cap but incomplete as yet emersion liquid level all moves into the containers that granulated glass sphere is housed, shake fragmentation, obtain to contain the suspension liquid of mycelia fragment.The bacteria suspension of mixing is inserted in the fermention medium by 15% inoculum size, cultivated 1-5 days, take by weighing mycelium over dry weight.The variation tendency of finding mycelium production is similar to Example 1, is higher than control medium (glucose) mycelium production equally, and is in ascendant trend, and 4～6 days is the growth animated period of mycelia.
The straw hydrolyzed solution is produced mycelium and is induced synthesis of oxalic acid decarboxylase (initial pH value of medium 5.0)
Appoint and to get embodiment 1～3 and cultivate 4～6 days mycelia, adding final concentration is the oxalic acid of 10mM, induce 0.5-5 days after, collect mycelium, preserve fermented liquid.Mycelium is used 0.2M after liquid nitrogen freezing grinds fragmentation, the acetate buffer solution of pH 3.7 extracts the mycelia fragment, centrifugal removal residue, and supernatant is crude enzyme liquid.Residue dries by the fire to over dry weighs, and supernatant is measured oxalate decarboxylase activity and the white content of enzyme liquid eggs, and fermented liquid is measured residual concentration of reduced sugar.
Control group adopts dextrose culture-medium, cultural method is identical, is cultivating Corilus versicolor Quel. after 4 days, and adding final concentration is the oxalic acid of 10mM, after inducing 1 to 5 day, collect mycelium, preserve fermented liquid, after hypha fluid chilled nitrogen grinding fragmentation, use 0.2M, the acetate buffer solution of pH 3.7 extracts the mycelia fragment, centrifugal removal residue, and supernatant is crude enzyme liquid.Residue dries by the fire to over dry weighs, and supernatant is measured oxalate decarboxylase activity and the white content of enzyme liquid eggs, and fermented liquid is measured residual concentration of reduced sugar.Conclusion:
(a) mycelium produces the comparison that the oxalate decarboxylase enzyme is lived: straw hydrolyzed solution group enzyme activity reached 1.3U at 24 hours, reached high some 2U at 120 hours, and control group reached 0.3U at 48 hours, well below straw hydrolyzed solution group (see figure 2).
(b) mycelium produces the comparison than vigor of protein content and oxalate decarboxylase albumen: the straw hydrolyzed solution is cultivated mycelium and is induced the back protein content along with the prolongation of time is fallen after rising, and arrives high some 0.57mg at 72 hours; The time dependent fluctuation of the protein content of control group is less, maintains 0.1mg left and right sides (see figure 3).The broken gained zymoprotein of mycelium vigor is got the enzyme liquid eggs in vain than work (being labeled as than vigor 1) divided by enzyme liquid protein content, and straw hydrolyzed solution substratum is higher at 24 hours than vigor, reaches 12U/mg, descends subsequently, rises slightly with alive rising of enzyme after 72 hours.Control group only reached higher point than vigor at 48 hours be the 5.7U/mg (see figure 4).As seen inducing 24 hours, is the more satisfactory mycelial time of results.
(c) the heavy comparison of residue over dry after the mycelium fragmentation: the straw hydrolyzed solution cultivate mycelium induce the broken mycelia in back filter residue, being baked to over dry under 105 ℃ weighs, the residue dry weight was being induced 48 hours, promptly cultivate after 8 days and tend to be steady, about the 0.12g mycelium, and control group is about the 0.5g mycelium, the later stage (see figure 5) that slightly rises.The broken gained zymoprotein of mycelium that fermented liquid produces vigor is got enzyme liquid dry weight than vigor (being labeled as than vigor 2) divided by mycelia residue dry weight, straw hydrolyzed solution substratum is more higher 24 hours and 120 hours than living, reach the 15U/g mycelium, roughly the same with enzyme activity trend.And control group reached high some 6U/g mycelium (see figure 6) at 48 hours.
(d) comparison of the residual reducing sugar content in straw substratum fermentation back: after the straw hydrolyzed solution substratum fermentation culture, reducing sugar content constantly reduces, and approaches zero after inducing 5 days.And control group sugar degree behind fermentation inducement maintains 16g/L left and right sides (see figure 7).Show that Corilus versicolor Quel. can make full use of the reducing sugar in the straw hydrolyzed solution.
(e) variation of straw substratum fermentation back pH value: the slow ascendant trend of pH value maintenance after the fermentation of straw substratum, from 24 hours to 120 hours, rise to 6.0 from 5.0.The pH value of control group was that production of enzyme reaches high point 3.4 when the highest at second day, then maintained 3 left and right sides (see figure 8)s.Illustrate that high pH value does not exert an influence to inducing Corilus versicolor Quel. to produce oxalate decarboxylase in straw hydrolyzed solution substratum.
(f) mycelium produces the comparison than vigor of protein content and oxalate decarboxylase albumen: the straw hydrolyzed solution is cultivated mycelium and is induced the back protein content along with the prolongation of time is fallen after rising, and arrives high some 0.57mg at 72 hours; The time dependent fluctuation of the protein content of control group is less, maintains 0.1mg left and right sides (see figure 3).The broken gained zymoprotein of mycelium vigor is got the enzyme liquid eggs in vain than work (being labeled as than vigor 1) divided by enzyme liquid protein content, and straw hydrolyzed solution substratum is higher at 24 hours than vigor, reaches 12U/mg, descends subsequently, rises slightly with alive rising of enzyme after 72 hours.Control group only reached higher point than vigor at 48 hours be the 5.7U/mg (see figure 4).As seen inducing 24 hours, is the more satisfactory mycelial time of results.
(g) the heavy comparison of residue over dry after the mycelium fragmentation: the straw hydrolyzed solution cultivate mycelium induce the broken mycelia in back filter residue, being baked to over dry under 105 ℃ weighs, the residue dry weight was being induced 48 hours, promptly cultivate after 8 days and tend to be steady, about the 0.12g mycelium, and control group is about the 0.5g mycelium, the later stage (see figure 5) that slightly rises.The broken gained zymoprotein of mycelium that fermented liquid produces vigor is got enzyme liquid dry weight than vigor (being labeled as than vigor 2) divided by mycelia residue dry weight, straw hydrolyzed solution substratum is more higher 24 hours and 120 hours than living, reach the 15U/g mycelium, roughly the same with enzyme activity trend.And control group reached high some 6U/g mycelium (see figure 6) at 48 hours.
(h) comparison of the residual reducing sugar content in straw substratum fermentation back: after the straw hydrolyzed solution substratum fermentation culture, reducing sugar content constantly reduces, and approaches zero after inducing 5 days.And control group sugar degree behind fermentation inducement maintains 16g/L left and right sides (see figure 7).Show that Corilus versicolor Quel. can make full use of the reducing sugar in the straw hydrolyzed solution.
(i) variation of straw substratum fermentation back pH value: the slow ascendant trend of pH value maintenance after the fermentation of straw substratum, from 24 hours to 120 hours, rise to 6.0 from 5.0.The pH value of control group was that production of enzyme reaches high point 3.4 when the highest at second day, then maintained 3 left and right sides (see figure 8)s.Illustrate that high pH value does not exert an influence to inducing Corilus versicolor Quel. to produce oxalate decarboxylase in straw hydrolyzed solution substratum.
The straw hydrolyzed solution is produced Corilus versicolor Quel. (initial pH value of medium 3.5)
The substratum compound method after preparation is finished, is regulated pH value to 3.5 with sodium hydroxide with embodiment 1, and the sterilization back is standby.
Inoculation and cultural method are with embodiment 1, and the hydrolyzed solution substratum mycelium production of initial pH 3.5 is very low, far below the mycelial biomass (see figure 9) of contrast dextrose culture-medium.
Claims (6)
1. a straw carbon source is cultivated the method for Corilus versicolor Quel. inducing oxalate decarboxylase, comprising:
(1) be 1g with the straw of pulverizing according to solid-to-liquid ratio: 5-15ml soaks 10-20h with acid solution, and hydrolysis reaction 20-60min under 100-150 ℃ of condition behind the centrifugal removal residue, obtains the straw hydrolyzed solution, and measures reducing sugar content in the hydrolyzed solution with the DNS method;
(2) press the reducing sugar amount of 7-30g/L, the straw hydrolyzed solution is mixed with fermention medium, regulating pH value be 4.0-5.0, removes by filter impurity, and adds nitrogenous source and inorganic salt, reexamines and regulate the pH value to 4.0-5.0, and it is afterwards standby to sterilize;
(3) getting 2-3 sheet diameter is that 9～11mm contains mycelial agar solid medium, inserts liquid seed culture medium, leaves standstill in 30 ℃ and cultivates 4-6 days; Treat that mycelium liquid medium within top forms the white hypha group but when not emerging as yet, mycelium and substratum are all transferred in the container that contains sterile glass beads, mycelium is shaken the broken suspension liquid that obtains to contain the mycelia fragment, inoculum size with 3-15% inserts above-mentioned fermention medium, cultivates 4-6 days under 20-30 ℃ and 100-250rpm speed conditions;
When (4) treating that mycelium pellet forms, adding final concentration is the generation of the oxalic acid inducing oxalate decarboxylase of 5-20mM, cultivates and obtains mycelium after 0.5-6 days; Mycelium grinds broken behind liquid nitrogen freezing, uses 0.2M, and the acetate buffer solution of pH3.7 extracts the mycelia fragment, centrifugal removal residue, and the gained supernatant liquor is the oxalate decarboxylase crude enzyme liquid.
2. a kind of straw carbon source according to claim 1 is cultivated the method for Corilus versicolor Quel. inducing oxalate decarboxylase, and it is characterized in that: the acid in the described step (1) is sulfuric acid, hydrochloric acid or acetic acid, and mass percent concentration is 0.5%-8%.
3. a kind of straw carbon source according to claim 1 is cultivated the method for Corilus versicolor Quel. inducing oxalate decarboxylase, it is characterized in that: described step (2) is regulated the pH value with sodium hydroxide solution.
4. a kind of straw carbon source according to claim 1 is cultivated the method for Corilus versicolor Quel. inducing oxalate decarboxylase, and it is characterized in that: the component of the fermention medium in the described step (2) is: the reducing sugar 7～30g in the straw hydrolyzate, peptone 3.0g, KH
2PO
41.0g, Na
2HPO
412H
2O 0.2g, MgSO
47H
2O 0.5g and micro-1ml, water is settled to 1L.
5. a kind of straw carbon source according to claim 4 is cultivated the method for Corilus versicolor Quel. inducing oxalate decarboxylase, and it is characterized in that: the component of described trace element is: FeSO
47H
2O 10g, MnSO
4H
2O 1.0g, ZnSO
47H
2O 1.0g, CuSO
45H
2O 2.0g, CaCl
22H
2O 13g, water is settled to 1L.
6. a kind of straw carbon source according to claim 1 is cultivated the method for Corilus versicolor Quel. inducing oxalate decarboxylase, it is characterized in that: the liquid seed culture medium in the described step (3), with the reducing sugar in the glucose replacement fermention medium, all the other components are identical with fermention medium.
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| CN102533707A (en) * | 2012-01-06 | 2012-07-04 | 东华大学 | Method of culturing coriolus versicolor-induced oxalate decarboxylase through jerusalem artichoke carbon source |
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| CN102533707A (en) * | 2012-01-06 | 2012-07-04 | 东华大学 | Method of culturing coriolus versicolor-induced oxalate decarboxylase through jerusalem artichoke carbon source |
| CN102533707B (en) * | 2012-01-06 | 2016-12-14 | 东华大学 | A kind of method of culturing coriolus versicolor-induced oxalate decarboxylase through jerusalem artichoke carbon source |
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