CN103087922A - Penicillium, cellulose produced by fermenting penicillium in solid state and method thereof for preparing pomelo peel high-ester pectin - Google Patents
Penicillium, cellulose produced by fermenting penicillium in solid state and method thereof for preparing pomelo peel high-ester pectin Download PDFInfo
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Abstract
The invention discloses penicillium, cellulose produced by fermenting penicillium in a solid state and a method thereof for preparing pomelo peel high-ester pectin. A penicillium strain is penicillium (Penicillium sp.) WZUX01 and is preserved in the General Micro-Organism Centre of the China Micro-Organism Strain Preservation and Management Committees; and the preservation number is CGMCC NO. 6370. Cellulose is produced by fermenting the penicillium (Penicillium sp.) WZUX01 in the solid state; the cellulose can be used for preparing pomelo peel high-ester pectin; and the content of the methoxy group in the pomelo peel pectin prepared by the method can be up to 30.48%.
Description
Technical field
The invention belongs to zymetology (C12P) technology, be specifically related to a penicillium, its solid state fermentation cellulase-producing, and this cellulase is used for the method for shaddock ped high ester pectin processed.
Background technology
Pectin (Pectin) is a kind of wetting ability vegetable jelly, extensively is present in the cell walls of root, stem, leaf, fruit of higher plant.The pectin main component is D-galacturonic acid (D-galactuonic acid), and wherein the part galacturonic acid is by esterification, and molecular weight is between 3～180,000.Commercialization pectin has liquid pectin and pectin powder, and the color and luster of pectin is from oyster white to khaki, because of raw material, production technique difference.According to gamma value (DM) difference, pectin is divided into high methoxyl pectin (gamma value is equivalent to methoxy content greater than 7% greater than 50%) and low-methoxy pectin (gamma value is equivalent to methoxy content less than 7% less than 50%), and the latter comprises acid amides pectin.
Pectin has good emulsifying, thickening, stable and pectisation, is widely used in fields such as food, weaving, printing and dyeing, tobacco, metallurgy.Because pectin has the effects such as antibiotic, hemostasis, detumescence, detoxifcation, antidiarrheal, reducing blood-fat, radioprotective, or a kind of good pharmaceutical preparation matrix, can be used to make laxative, hemostatic agent, toxic metal toxinicide, plasma substitute etc., is medicine and the indispensable auxiliary material of cosmetic industry.Pectin is the main component of water-soluble dietary fibre in the human body seven major nutrient, development research along with functional polysaccharide, pectin is as water-soluble dietary fibre, more and more is subject to studying the attention with processing industry, has the multiple effects such as good diarrhea, anticancer, treatment diabetes and fat-reducing.
Shaddock is the mature fruit of rue family citrus plant evergreen fruit trees, plants in a large number in south China many areas.China's shaddock ped aboundresources, but major part is dropped, and caused environmental pollution.In shaddock ped, pectin content is higher, and contained pectin degree of gelation is high, rank is high.Therefore, extract pectin from shaddock ped and have higher economic benefit and social benefit.
Extract pectin existing commercialization production at present from shaddock ped, many bibliographical informations are also arranged, reported that as Zhou Jinhua the acid system that obtains by orthogonal experiment extracts the processing condition of pomelo-pectin, pectin yield is the 14.9%(research and development of natural products, 2006,18:483-486), and for example Zhao Mei etc. has reported and has adopted acid hydrolyzation to extract the processing condition of shaddock ped low-methoxy pectin, pectin yield is the 5.51%(food and pharmaceutical, 2008,10(7): 29-31).
Although acid extraction method is still the most frequently used pectin extraction method at present, acid extraction easily causes environmental pollution.Therefore people actively inquire into other extracting method of environmental contamination reduction.As ion exchange method (Sun Run, foodstuffs industry science and technology, 1989, (2): 40-41), microwave extraction method (Dredycfuss M S, Appl Eneerio Nicobio, 1980,38 (1): 13; M Kratchanova, Carbohydrate Polymers, 2004,56:181～185; Kong Zhen, Journal of Zhengzhou Grain College, 2000,21(2): 11-15), these methods all are difficult to realize industrial applications.
The separation method of the most frequently used pectin is ethanol precipitation.Other separation methods also have report, as salting-out process (Deng Hong, Food science, 2002,23 (3): 57-60; Zhang Linghua, pesticide herd product exploitation, 2000, (11): 12～14).Pectin lighter color, the ash oontent that ethanol precipitation is produced is few, degree of gelation is high, quality better.Although the ethanol usage quantity is larger, if waste ethanol is reclaimed and recycle, can reduce production costs.
Summary of the invention
The technical problem to be solved in the present invention is to adopt the Enzymatic Extraction pomelo-pectin, extracts to alleviate acid system the environmental pollution that is caused, and extraction yield and acid system approach and even surpass acid system simultaneously.
In order to solve the problems of the technologies described above, the invention provides a penicillium solid state fermentation cellulase-producing and for the preparation of the novel method of shaddock ped high ester pectin.Its principal feature extracts the cellulase in solid-state culture for to carry out solid state fermentation with a penicillium, obtained by freeze drying cellulase powder.Extract pomelo-pectin with the cellulase powder that makes, with the pectin of ethanol precipitation precipitation zyme extract, will precipitate drying after centrifugal and make the pectin finished product, its methoxy content belongs to high ester pectin after measured.The shaddock ped high ester pectin preparation method that this invention provides, the pectin extraction rate of its pectin powder be higher than the present acid extraction method of report, and alleviate acid to the pollution of environment.
The invention provides a penicillium, this bacterial strain be Penicillium notatum (
Penicillium sp.) WZUX01, being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC NO. 6370.
The present invention also provides the method for above-mentioned Penicillium notatum solid state fermentation cellulase-producing, comprises the steps:
1) described bacterial strain Penicillium notatum WZUX01 line is transferred on the PDA slant medium, cultivates 3 ~ 6 days under 25 ~ 35 ℃;
2) Penicillium notatum in the culture that obtains with the water elution step 1) obtains spore suspension, and the straw bran mass of access sterilization was cultivated 5 ~ 7 days under 25 ~ 35 ℃, and wherein the straw bran mass is made of the composition of following weight ratio: rice straw powder: wheat bran: (NH
4)
2SO
4: water=4 ~ 36:8 ~ 48: 0 ~ 16: 24 ~ 80;
3) the solid-state culture that step 2) obtains adds distilled water to smash to pieces, and 35 ~ 45 ℃ of extracting at constant temperature 2 ~ 10h filter, and get enzyme liquid, namely get cellulase through lyophilize.Preferably, after above-mentioned extracting at constant temperature 2 ~ 10h, be placed in and soak 24 h under 4 ℃.
Preferably, above-mentioned steps 2) described in the spore amount of spore suspension be 1 * 10
12~ 9 * 10
12Individual/mL, the inoculum size of described spore suspension: the envelope-bulk to weight ratio of spore suspension and straw bran mass is 1 mL: 1 ~ 3 g is preferably 1:2.
Preferably, step 2) described straw bran mass is made of the composition of following weight ratio: rice straw powder: wheat bran: (NH
4)
2SO
4: water=8:32:8:52.
The cellulase that the method for above-mentioned Penicillium notatum solid state fermentation cellulase-producing obtains.
The present invention also provides the above-mentioned cellulase method for the preparation of the shaddock ped high ester pectin, comprises the steps:
1) with shaddock ped oven dry, pulverizing, get the shaddock ped powder;
2) the shaddock ped powder is added in enzyme reactor, add damping fluid, then add described cellulase to extract, then the centrifugal liquid pectin that gets;
3) regulating step 2) pH of gained liquid pectin is 5 ~ 9, adds ethanol, precipitation, centrifugal throw out, drying gets pectin.Sedimentation time is preferably 2 ~ 32h.
Preferably, step 2) described in, the pH value of damping fluid is 5 ~ 7, and the weightmeasurement ratio of shaddock ped powder and described damping fluid is 5 ~ 20 g:100 mL, and described damping fluid more preferably pH value is 7 HAC/NaAC damping fluid.
The weight that adds of cellulase preferably, step 2) is 1% ~ 3% of described shaddock ped grain weight amount.
Preferably, the condition of extracting step 2) is: extract 24 ~ 48 h under 35 ~ 45 ℃.
Preferably, in step 3), regulating step 2) pH of gained liquid pectin is 5 ~ 9, the concentration of volume percent in liquid pectin is 60 ~ 80% to it to add ethanol.
The present invention can reach following effect: Penicillium notatum (
Penicillium sp.) ability of WZUX01 solid state fermentation cellulase-producing is that the solid-state cultivation produce of every gram CMC enzyme activity is 8210.667U, the CMC enzyme activity of prepared cellulase powder is 300U/mg; The shaddock ped Steamed dumpling with pork，mushrooms and bamboo shoots glue extraction yield that the cellulase powder extracts pomelo-pectin is 19.30%, the pectin deposition rate of ethanol precipitated pectin is 80.23%, the pectin finished product extraction yield of shaddock ped powder is 15.48%, and the methoxy content of gained pectin finished product can reach 30.48%, belongs to high ester pectin.
Description of drawings
Fig. 1 be Penicillium notatum (
Penicillium sp.) WZUX01 Produced by Solid-state Fermentation cellulase curve.
Fig. 2 is that temperature is to cellulase powder CMC enzyme activity influence curve.
Fig. 3 is that pH is to cellulase powder CMC enzyme activity influence curve.
The bacterial strain preservation
Mould of the present invention (
Penicillium sp.) WZUX01, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, the preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, deposit number are CGMCC NO. 6370, and preservation date is on July 20th, 2012.
Embodiment
The invention will be further described below in conjunction with the drawings and specific embodiments so that those skilled in the art the present invention may be better understood and can be implemented, but illustrated embodiment is not as a limitation of the invention.
Embodiment one: Penicillium notatum (
Penicillium sp.) WZUX01 solid state fermentation cellulase-producing
Bacterial strain
Penicillium notatum (
Penicillium sp.) WZUX01, separate acquisition with environmental science institute by Wenzhou University's life in the rice straw compost soil of tea hill, Wenzhou, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC NO. 6370.
(2) substratum
Potato substratum (PDA substratum): murphy juice 1000mL, agar 20g, pH nature.The preparation of murphy juice: get the fresh peeled potatoes of 200g, be cut into small pieces, after adding 1000mL distillation poach 1h, with filtered through gauze, mend to 1000mL with distilled water.
The weight ratio of each component of straw bran mass is: rice straw powder: wheat bran: (NH
4)
2SO
4: water=4 ~ 36:8 ~ 48:0 ~ 16:24 ~ 80, pH nature.
(3) method
Preservation strain Penicillium notatum WZUX01 line is transferred on the PDA slant medium, cultivated 4 days for 30 ℃, the sterilized water of 10mL is added in the test tube that has covered with mould several times, vibration makes spore suspension, measure the spore quantity of spore suspension, spore suspension is accessed sterilized straw bran mass, stir, cultivated 4 days under 25-35 ℃, every day, the stir culture base was 1 to 2 time.Take the solid-state culture 10g that obtains, add the distilled water of 90mL to smash to pieces, vibration lixiviate 4h in 40 ℃, 150r/min water bath with thermostatic control, be placed in refrigerator (4 ℃) and soak 24 h, filtered through gauze gets crude enzyme liquid, centrifugal 15 minutes kinds under 4000rpm, get supernatant liquor (enzyme liquid), measure the CMC enzyme activity.
CMC enzyme activity determination method: the enzyme liquid 0.5mL after absorption is suitably diluted adds the citric acid-sodium citrate damping fluid (0.05molL that contains 0.5%CMC-Na in test tube
-1, then pH4.4) 1.5 mL after reacting 30 min in 50 ℃ of water-baths, often in vitro add 1.5mLDNS reagent immediately, then boil 5 min in boiling water after, using immediately cold water cooling, be settled to 25mL, in the 520nm mensuration OD of place value, try to achieve its reducing sugar content from typical curve.The CMC enzyme activity unit is defined as the enzyme amount of hydrocellulose generation 1 μ g reducing sugar amount in per minute.The enzyme liquid of deactivation 10min is blank (Wu Hailong etc., science and technology circular, 2008,24 (6): 769～776) in 100 ℃ of water-baths.
The spore quantity of spore suspension adopts the blood counting chamber method to measure.
The setting of straw bran mass formula and culture condition and the results are shown in following table 1.
Each experimental group straw bran mass total amount is 20g, and the quantity of spore suspension is 1 * 10
12～9 * 10
12/ mL.
Setting and the result of table 1 straw bran mass formula and culture condition
| Experimental group | Straw: wheat bran: (NH 4) 2SO 4: water (W:W:W:W) | Temperature (℃) | CMC enzyme activity (U/g) |
| 1 | 36:24:0:40 | 25 | 3960.6665 |
| 2 | 16:24:16:44 | 25 | 2390.8890 |
| 3 | 4:16:8:72 | 25 | 3140.5000 |
| 4 | 24:36:0:40 | 30 | 5270.9445 |
| 5 | 8:32:8:52 | 30 | 7220.500 |
| 6 | 12:8:16:64 | 30 | 3840.3890 |
| 7 | 12:48:16:24 | 35 | 2120.5000 |
| 8 | 24:16:8:52 | 35 | 2000.2220 |
| 9 | 8:12:0:80 | 35 | 4590.000 |
In table 1, temperature refers to the culture temperature of the sterilized straw bran mass of spore suspension access, and the CMC enzyme activity refers to the CMC enzyme activity of the solid-state culture of every g.
The CMC enzyme activity data of each experimental group can be found out from table 1, and the best proportioning of straw bran mass is experimental group 5, i.e. rice straw powder: wheat bran: (NH
4)
2SO
4: water=8: 32: 8: 52, pH nature.As take rice straw powder 1.60g, wheat bran 6.40g, (NH
4)
2SO
41.6g in 250 mL beakers, add 10.4 mL distilled water, stir with glass stick.
Produce the enzyme curve determination under optimal conditions, result as shown in Figure 1.It produces the enzyme peak value about 144 h as can be known by accompanying drawing 1, namely cultivates 6 days, and the CMC enzyme activity can reach 8210.667U/g.
Embodiment two: the characteristic measurement of cellulase powder
(1) preparation of cellulase powder
With the Penicillium notatum of preservation (
Penicillium sp.) WZUX01 transfers in the PDA slant medium, cultivates 4 days under 30 ℃, obtains the straw bran mass of filling a prescription by embodiment one experimental group 5 of spore suspension access sterilization with the sterilized water wash-out, the spore amount of spore suspension is 1 * 10
12~ 9 * 10
12/ mL, the inoculum size of spore suspension is spore suspension: straw bran mass=1:2 (V:W, mL/g), after stirring, cultivated 6 days under 30 ℃, add distilled water to smash to pieces, vibration lixiviate 4h in 40 ℃, 150r/min water bath with thermostatic control, be placed in and soak 24 h under 4 ℃ of low temperature, filter to get crude enzyme liquid, get the cellulase powder through lyophilize.
(2) cellulase powder optimum temperuture is measured
Take appropriate cellulase powder 0.05molL
-1, pH4.4 the citric acid-sodium citrate damping fluid be mixed with enzyme liquid, press embodiment one method and measure respectively the CMC enzyme activity under 25 ℃, 30 ℃, 35 ℃, 40 ℃, 45 ℃, 50 ℃, 55 ℃ and 60 ℃, the results are shown in Figure 2.As shown in Figure 2, the optimum temperuture of cellulase powder is 35 ~ 45 ℃.
In Fig. 2, the highest CMC enzyme activity of the CMC enzyme activity that records at enzyme activity=a certain temperature/record * 100%.
(3) cellulase powder optimal pH is measured
Respectively take appropriate cellulase powder and be respectively 3.0,4.0,5.0,6.0,7.0,8.0,9.0 damping fluid with pH and be mixed with enzyme solution, wherein pH 3.0,4.0,5.0 and 6.0 uses 0.05molL
-1The citric acid-sodium citrate damping fluid, pH is 7.0,8.0 and 9.0 the 0.05molL that uses
-1The HAC/NaAc damping fluid, use from prepare cellulase powder solution phase with damping fluid prepare respectively the 0.5% CMC-Na solution of different pH, the CMC enzyme activity by under the different pH of embodiment one method (measuring temperature is 40 ℃) mensuration the results are shown in Figure 3.As shown in Figure 3, the optimal pH of cellulase powder is 5 ~ 7.In Fig. 3, the highest CMC enzyme activity of the CMC enzyme activity that records under enzyme activity=a certain pH/record * 100%.
(4) mensuration of cellulase powder CMC enzyme activity
Take appropriate cellulase powder 0.05molL
-1, pH 6.0 the citric acid-sodium citrate damping fluid be mixed with enzyme liquid, measure the CMC enzyme activity by embodiment one method (measure temperature be 40 ℃), the CMC enzyme activity of cellulase powder is 300U/mg after measured.
Embodiment three: the cellulase powder extracts pomelo-pectin
(1) shaddock ped is processed
The fresh pomelo peel of Yongjia County, Zhejiang Province is originated from purchase, is cut into fine grained chippings with scissors and is placed in baking oven dries (24-72 h) under 60 ℃, pulverizes, and gets the shaddock ped powder and preserves with standby.
(2) method
Take a certain amount of shaddock ped powder in enzyme reactor, the HAC/NaAC damping fluid that adds pH 7.0, the shaddock ped powder: damping fluid is 5 ~ 20:100(W:V, g/mL), add again 1% ~ 3% cellulase powder (accounting for the weight percentage of shaddock ped powder) that makes by embodiment two (1) methods, extract 24 ~ 48 h under 35 ~ 45 ℃, then measure supernatant liquor (liquid pectin) concentration of pectin after centrifugal 10min under 4000rpm, calculate the pectin extraction rate of pectin powder.
The mensuration of concentration of pectin: get the 0.4mL pectin extracting soln and be settled to 50mL, put 4 test tubes in the beaker that is added with ice cube and water, each slowly adds the 6 mL vitriol oils, cooling limit, limit adds the liquid pectin 1mL of constant volume, wherein 1 adds distilled water to compare, and boils 10min after shaking up in boiling water, respectively adds 1mL carbazole ethanol after being cooled to room temperature, place 30min under room temperature, survey the OD value under 530nm, try to achieve concentration of pectin (Gu Fuchang, Food science from typical curve, 1984, (8): 45～46).
Pectin extraction rate (%)=supernatant liquor concentration of pectin (g/mL) * supernatant liquor volume (mL)/pectin powder amount (g) * 100%.
Following table 2 is set and be the results are shown in to the experiment condition of cellulase powder extracting pomelo-pectin.
The experiment condition of table 2 cellulase powder extracting pomelo-pectin is set and result
| Experimental group | Shaddock ped powder: damping fluid (W:V is as g:mL) | The cellulase powder (accounts for shaddock ped powder percentage ratio, %) | Extraction time (h) | The extraction temperature (℃) | Pectin extraction rate (%) |
| 1 | 5:100 | 1 | 24 | 35 | 6.30 |
| 2 | 5:100 | 2 | 36 | 40 | 10.78 |
| 3 | 5:100 | 3 | 48 | 45 | 7.68 |
| 4 | 10:100 | 1 | 48 | 45 | 16.74 |
| 5 | 10:100 | 2 | 36 | 40 | 19.30 |
| 6 | 10:100 | 3 | 24 | 35 | 14.32 |
| 7 | 20:100 | 1 | 48 | 40 | 6.86 |
| 8 | 20:100 | 2 | 24 | 45 | 9.66 |
| 9 | 20:100 | 3 | 36 | 35 | 5.52 |
The pectin extraction rate of experimental group 5 pectin powders is the highest as shown in Table 2, reaches 19.30%.
Embodiment four: the ethanol precipitation precipitated pectin
Prepare pomelo-pectin liquid by embodiment three experimental group 5 methods, respectively getting liquid pectin 100mL adjusting pH is 5 ~ 9, then the concentration of volume percent in liquid pectin reaches 60 ~ 80% to it to add ethanol, precipitation 2 ~ 32 h, get after centrifugal 10min after precipitation finishes to be deposited under 4000rpm and be dried to constant weight under 90 ~ 100 ℃ and get dry pectin, weighing pectin weight is calculated pectin precipitation yield.
Pectin amount (g)/liquid pectin concentration of pectin (g/mL) * liquid pectin volume (mL) * 100 after precipitation yield (%)=drying.
The experiment condition of ethanol precipitation precipitated pectin and the results are shown in following table 3.
Experiment condition and the result of table 3 ethanol precipitation precipitated pectin
| Experimental group | Precipitation pH | Alcohol concn (%) | Sedimentation time (h) | Pectin deposition rate (%) |
| 1 | 5 | 60 | 2 | 51.80 |
| 2 | 5 | 70 | 8 | 58.85 |
| 3 | 5 | 80 | 32 | 44.01 |
| 4 | 7 | 60 | 32 | 73.29 |
| 5 | 7 | 70 | 8 | 80.23 |
| 6 | 7 | 80 | 2 | 62.84 |
| 7 | 9 | 60 | 32 | 15.20 |
| 8 | 9 | 70 | 2 | 17.51 |
| 9 | 9 | 80 | 8 | 14.63 |
As shown in Table 3, the precipitation yield of experimental group 5 pectin is the highest, reaches 80.23%.
Embodiment five: the mensuration of pectin product methoxyl group
The measuring method of pectin product methoxyl group: accurately take the pectin finished product of 0.6-1.0g with analytical balance, after adding 100mL distilled water and dissolving fully, add 2 of 1% phenolphthalein indicators, use 0.1molL
-1NaOH is titrated in pink 0.5min colour-fast for terminal point, then adds 20mL 0.5 molL
-1The NaOH standardized solution, saponification 2.5 h use 0.5molL
-1H
2SO
4Standardized solution is titrated to red the disappearance, calculates pectin methoxy content (Gu Fuchang, Food science, 1984, (8): 45～46).
Pectin methoxy content (%)=[(M
1V
1-MV) * 0.031]/W * 100
V
1: the volume (mL) of the NaOH standardized solution that saponification adds
M
1: the volumetric molar concentration (molL of the NaOH standardized solution that saponification adds
-1)
V: the H that the alkali of titration saponification amount consumes
2SO
4The volume of standardized solution (mL)
M:H
2SO
4Volumetric molar concentration (the molL of standardized solution
-1)
0.031: the mmole number of methoxyl group
W: the quality of pectin finished product (g)
After measured, by the pectin that method of the present invention obtains, methoxy content is 30% left and right, belongs to high ester pectin.
And according to top condition: take 3 parts of liquid pectins that extract by embodiment three experimental group 5 use cellulase powder and through by the example four experimental group 5 dried pectin finished products of ethanol precipitation, measure its methoxy content, the methoxyl group average content that records pectin is 30.48%.
The above embodiment is the preferred embodiment that proves absolutely that the present invention lifts, and protection scope of the present invention is not limited to this.Being equal to that those skilled in the art do on basis of the present invention substitutes or conversion, all within protection scope of the present invention.Protection scope of the present invention is as the criterion with claims.
Claims (10)
1. a penicillium, is characterized in that, this bacterial strain be Penicillium notatum (
Penicillium sp.) WZUX01, being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC NO. 6370.
2. the method for Penicillium notatum solid state fermentation cellulase-producing claimed in claim 1, is characterized in that, comprises the steps:
1) described bacterial strain Penicillium notatum WZUX01 line is transferred on the PDA slant medium, cultivates 3 ~ 6 days under 25 ~ 35 ℃;
2) Penicillium notatum in the culture that obtains with the water elution step 1) obtains spore suspension, and the straw bran mass of access sterilization was cultivated 5 ~ 7 days under 25 ~ 35 ℃, and wherein the straw bran mass is made of the composition of following weight ratio: rice straw powder: wheat bran: (NH
4)
2SO
4: water=4 ~ 36:8 ~ 48: 0 ~ 16: 24 ~ 80;
3) the solid-state culture that step 2) obtains adds distilled water to smash to pieces, and 35 ~ 45 ℃ of extracting at constant temperature 2 ~ 10h filter to get enzyme liquid, get cellulase through lyophilize.
3. method according to claim 2, is characterized in that step 2) described in the spore amount of spore suspension be 1 * 10
12~ 9 * 10
12Individual/mL, the inoculum size of described spore suspension: the envelope-bulk to weight ratio of spore suspension and straw bran mass is 1mL:1～3g.
4. method according to claim 2, is characterized in that step 2) described straw bran mass is made of the composition of following weight ratio: rice straw powder: wheat bran: (NH
4)
2SO
4: water=8:32:8:52.
5. the cellulase that obtains of the method for the described Penicillium notatum solid state fermentation of claim 2 ~ 4 any one cellulase-producing.
6. cellulase claimed in claim 5 for the preparation of the method for shaddock ped high ester pectin, is characterized in that, comprises the steps:
1) with shaddock ped oven dry, pulverizing, get the shaddock ped powder;
2) the shaddock ped powder is added in enzyme reactor, add damping fluid, then add described cellulase to extract, then the centrifugal liquid pectin that gets;
3) regulating step 2) pH of gained liquid pectin is 5 ~ 9, adds ethanol, precipitation, centrifugal, drying precipitate gets pectin.
7. method according to claim 6, is characterized in that step 2) described in the pH value of damping fluid be 5 ~ 7, the weightmeasurement ratio of shaddock ped powder and described damping fluid is 5 ~ 20g:100mL.
8. method according to claim 6, is characterized in that step 2) described in the weight that adds of cellulase be 1% ~ 3% of described shaddock ped grain weight amount.
9. method according to claim 6, is characterized in that step 2) in the condition extracted be: extract 24 ~ 48 h under 35 ~ 45 ℃.
10. method according to claim 6, is characterized in that, in step 3), regulating step 2) pH of gained liquid pectin is 5 ~ 9, the concentration of volume percent in liquid pectin is 60 ~ 80% to it to add ethanol.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103720733A (en) * | 2014-01-23 | 2014-04-16 | 程钰翔 | Preparation method of manyprickle acanthopanax root extract |
| CN115125152A (en) * | 2022-03-28 | 2022-09-30 | 湖南科技学院 | Mixed bacteria for degrading lignocellulose, mixed enzyme and degradation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5877014A (en) * | 1997-05-29 | 1999-03-02 | University Of Massachusetts | Penicillium strain for bioremediation |
| CN101717728A (en) * | 2009-12-28 | 2010-06-02 | 华东理工大学 | Penicillium and application thereof in catalyzing and hydrolyzing lignocellulose |
-
2012
- 2012-10-29 CN CN201210420183.3A patent/CN103087922B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5877014A (en) * | 1997-05-29 | 1999-03-02 | University Of Massachusetts | Penicillium strain for bioremediation |
| CN101717728A (en) * | 2009-12-28 | 2010-06-02 | 华东理工大学 | Penicillium and application thereof in catalyzing and hydrolyzing lignocellulose |
Non-Patent Citations (1)
| Title |
|---|
| 胡丹，等: "高产纤维素酶青霉菌的筛选及产酶条件的研究", 《中国食品添加剂》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103720733A (en) * | 2014-01-23 | 2014-04-16 | 程钰翔 | Preparation method of manyprickle acanthopanax root extract |
| CN115125152A (en) * | 2022-03-28 | 2022-09-30 | 湖南科技学院 | Mixed bacteria for degrading lignocellulose, mixed enzyme and degradation method |
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| CN103087922B (en) | 2014-05-28 |
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