CN108753492B - Application of chaetomium thermophilum cellulase in preparation of detergent - Google Patents
Application of chaetomium thermophilum cellulase in preparation of detergent Download PDFInfo
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- CN108753492B CN108753492B CN201810722622.3A CN201810722622A CN108753492B CN 108753492 B CN108753492 B CN 108753492B CN 201810722622 A CN201810722622 A CN 201810722622A CN 108753492 B CN108753492 B CN 108753492B
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- cellulase
- chaetomium
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- cotton
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38645—Preparations containing enzymes, e.g. protease or amylase containing cellulase
-
- C—CHEMISTRY; METALLURGY
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
-
- C—CHEMISTRY; METALLURGY
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
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- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M16/00—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic
- D06M16/003—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic with enzymes or microorganisms
-
- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M2101/00—Chemical constitution of the fibres, threads, yarns, fabrics or fibrous goods made from such materials, to be treated
- D06M2101/02—Natural fibres, other than mineral fibres
- D06M2101/04—Vegetal fibres
- D06M2101/06—Vegetal fibres cellulosic
Abstract
The invention discloses an application of a Chaetomium thermophilum cellulase in preparation of a detergent, wherein the Chaetomium thermophilum cellulase is expressed by trichoderma reesei. The thermophilic chaetomium cellulase has a good removal effect on cotton fabric hairiness within the pH value range of 8.0-11.0, and the removal effect on the cotton fabric hairiness is relatively small along with the fluctuation of the pH value; after the cotton or cotton blended fabric is washed for multiple times by using the detergent added with the chaetomium thermophilum cellulose, the hairiness on the surface of the cotton or cotton blended fabric can be effectively removed, the texture of the fabric is cleaned, the fabric is bright in luster and bright in color, and the effect of enabling the fabric to be durably renewed is achieved.
Description
Technical Field
The invention relates to the field of biological enzyme preparations, in particular to a new application of Chaetomium thermophilum cellulase, and especially relates to an application of Chaetomium thermophilum cellulase in preparation of a detergent.
Background
The cotton clothes have good wearability such as nature, comfortable, air permeability, and the like, and are deeply loved by people. However, cotton clothes are fluffy after being worn for a long time or washed for many times, the specular reflection of light is weakened, the brightness of the fabric is reduced, and the fabric looks older; meanwhile, the color of the clothes is gradually dull due to the continuous deposition of stubborn stains on the surfaces of the clothes. The cellulase can mildly decompose cotton fibers, so that fluff on the surface of the cotton fabrics can be removed, and the cotton fibers are fluffy, thereby being beneficial to removing stubborn stains, recovering the beautiful color of the fabrics and achieving the effect of renewing.
After cotton fabrics, particularly cotton clothes, become old, brightness and vividness of the cotton fabrics can be increased by adding a brightening agent during washing or spraying the brightening agent after washing, so that a 'renewing' effect is achieved, but the added brightening agent has adverse effects on the wearing comfort performance of the clothes; the added brightener also has different degrees of harm to human bodies, in addition, the refreshing effect is temporary, and after washing, the brightener is partially or completely washed away, so that the refreshing effect is poor.
The cellulase is added into the detergent, the removal effect of the cellulase on cotton fabrics is utilized, and after the cotton or cotton blended fabrics are washed for multiple times by using the detergent added with the cellulose, hairiness on the surfaces of the cotton or cotton blended fabrics can be effectively removed, so that the problem of fabric aging is fundamentally solved, and the effect of enabling the fabrics to be durably 'renewed' is achieved. The method is simple and convenient to use, and the wearability and the safety performance of the clothes are not influenced.
The washing process of clothes is carried out under alkaline conditions, generally in the range of pH 8.0 to 11.0, which requires cellulase used for detergent to have stable and mild removal effect of lint of cotton fabric under the above pH conditions. Under the conditions, the cellulase in the current market has a large fluctuation of the removal effect of cotton fabric hairiness along with the change of pH value; in addition, the fluff removal effect on cotton fabrics is poor, and in order to solve the problems, the dosage of cellulase is generally increased, so that the waste of enzyme preparations is caused, and the cost of the cellulase-added detergent is also increased.
Disclosure of Invention
The invention aims to solve the technical problem of providing a new application of Chaetomium thermophilum cellulase, namely an application in preparing a detergent. On one hand, the problems of non-persistence and non-environmental protection in the 'renewing' finishing of cotton fabrics and cotton clothes by using the brightener are solved. In addition, the problem that the existing cellulase has poor effect of removing the hairiness on the surface of the cotton fabric within the pH value range of 8.0-11.0 is solved. In addition, the problem that the removal effect of the cellulase on the surface hairiness of the cotton fabric greatly fluctuates along with the change of the pH value within the range of 8.0-11.0 at present is solved.
The Chaetomium thermophilum cellulase related by the invention is the Chaetomium thermophilum cellulase with the patent application number of 201310280267.6, which is applied by the applicant at 7, 5.2013 and is named "The invention discloses thermophilic chaetomium cellulase and a preparation method and application thereof. Chaetomium thermophilum (Chaetomicum thermophilum) The genome library has a homologous protein, the function is unknown, and the gene sequence, the cDNA sequence and the protein sequence are respectively shown as SEQ ID NO.1, SEQ ID number 2 and SEQ ID number 3.
In order to solve the technical problems, the invention adopts the following technical scheme:
the application of the chaetomium thermophilum cellulase in the preparation of the detergent is characterized in that the chaetomium thermophilum cellulase is prepared according to the following method:
1) synthesizing a chaetomium thermophilum cellulase gene, wherein the sequence of the synthesized chaetomium thermophilum cellulase gene is shown in SEQ ID NO.2, and the synthesized gene code is more in line with the gene code of the high-efficiency expression of the trichoderma reesei;
2) constructing thermophilic chaetomium cellulase expression plasmid;
3) transforming trichoderma reesei and screening a thermophilic chaetomium cellulase high-producing strain to obtain the thermophilic chaetomium cellulase, wherein the amino acid sequence of the thermophilic chaetomium cellulase is shown as SEQ ID NO.3, and the thermophilic chaetomium cellulase secreted outside cells is the amino acid sequence of the 22 th to 314 th sites of the sequence shown as SEQ ID NO. 3; the step 3) is specifically as follows: first, byEcoRIDegrading the plasmid constructed in the step 2) to form a plasmid fragment, and transferring the plasmid fragment into a Trichoderma reesei RUT C-30 host cell by adopting a chemical conversion method; then culturing on a PDA culture medium containing hygromycin B, inoculating colonies which grow vigorously on the PDA culture medium, transferring spores to a fresh PDA culture medium through growth and germination to separate and purify transformants which are stable in transformation, inoculating the spores of the transformants to a trichoderma cellulase induction culture solution, after culturing, sampling and centrifuging, and taking supernatant to perform SDS-PAGE electrophoretic analysis; the pH value of the trichoderma cellulase induction culture solution is 4.8, and lactose is used as an inducer.
As the preferable technical scheme of the invention, the thermophilic chaetomium cellulase has good capability of degrading the surface hairiness of the cotton fabric within the pH value range of 8.0-11.0, and has good effect of removing the surface hairiness of the cotton fabric.
As a preferable technical scheme, the thermophilic chaetomium cellulase has relatively small enzyme activity fluctuation within the pH value range of 8.0-11.0, the enzyme activity fluctuation is within 30%, and the removal effect on the surface hairiness of cotton fabrics is stable.
As a preferable technical scheme of the invention, the detergent contains thermophilic Chaetomium cellulase, and the thermophilic Chaetomium cellulase accounts for the following mass percentage ranges in the detergent: 0.1-2.0%, preferably 0.3-1.0%.
As a preferable technical scheme of the invention, the thermophilic chaetomium cellulase has a suitable action temperature of 15-50 ℃ when the detergent is used, and the optimal action temperature is as follows: 20-40 ℃.
As a preferable technical scheme of the invention, the detergent has the function of improving the washfastness of the fabric after refreshing and finishing.
As a preferable technical scheme of the invention, the washing time of the detergent is 30-600 min.
As a preferred technical scheme of the invention, the dosage of the detergent ranges from 0.5 to 3.0 g/L.
Compared with the prior art, the invention has the beneficial effects that:
1. the thermophilic chaetomium cellulase has a good removal effect on cotton fabric hairiness within the pH value range of 8.0-11.0, and the removal effect on the cotton fabric hairiness is relatively small along with the fluctuation of the pH value; after the cotton or cotton blended fabric is washed for multiple times by using the detergent added with the chaetomium thermophilum cellulose, the hairiness on the surface of the cotton or cotton blended fabric can be effectively removed, the texture of the fabric is cleaned, the fabric is bright in luster and bright in color, and the effect of enabling the fabric to be durably renewed is achieved. Experiments prove that compared with the 'renewing' finishing of cotton fabrics by using brightening agents, the 'renewing' finishing of the cotton fabrics by using the detergent added with the thermophilic chaetomium cellulase is not only environment-friendly, but also has lasting effect.
2. Experiments prove that the thermophilic chaetomium cellulase has a good effect of removing hairiness on the surface of cotton fabrics within the pH value range of 8.0-11.0, the effect is 750 times that of cellulase YL400E, and the cost of the enzyme-added detergent can be greatly reduced.
3. Experiments prove that the thermophilic chaetomium cellulase has relatively small enzyme activity fluctuation within the pH value range of 8.0-11.0, the enzyme activity fluctuation is within 30 percent, and the effect of removing the cotton fabric surface hairiness is stable.
Detailed Description
The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention. The experimental procedures, for which specific conditions are not indicated in the examples, are generally carried out according to conventional conditions, for example as described in Sambrook et al, molecular cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions as recommended by the manufacturer.
Example 1 preparation of Thermomyces thermophilus cellulase expressed by Trichoderma reesei (see example 1 of the specification of the invention patent application publication No. 201310280267.6 CN 103343111A).
Example 2 cellulase pH profiling:
the Chaetomium thermophilum cellulase of the invention is analyzed for the filter paper activity at 50 ℃, and the test result is as follows:
TABLE 1
| pH | 8.0 | 8.5 | 9.0 | 9.5 | 10.0 | 10.5 | 11.0 |
| Vitality of the body | 618 | 563 | 500 | 503 | 458 | 435 | 433 |
| Relative Activity/%) | 100% | 91.1% | 80.9% | 81.4% | 74.1% | 70.4% | 70.1% |
From the activity analysis in table 1, the activity of the thermophilic chaetomium cellulase is maintained to be more than 70% within the range of pH value of 8.0-11.0, the activity fluctuation of the cellulase is within 30%, and the activity is relatively stable.
The commercial cellulase YL400E (available from Guangzhou, Atmpound Biotech, Inc.) was analyzed for filter paper viability at 50 ℃ and the results were as follows:
TABLE 2
| pH | 8.0 | 8.5 | 9.0 | 9.5 | 10.0 | 10.5 | 11.0 |
| Vitality of the body | 782 | 580 | 402 | 245 | 159 | 146 | 133 |
| Relative Activity/%) | 100% | 74.2% | 51.4% | 31.3% | 20.3% | 18.7% | 17.0% |
From the activity analysis in table 2, the activity change of the cellulase in the market is large in the range of the pH value of 8.0-10.5, the activity of the cellulase at the pH values of 10.5 and 11.0 is about 20% of that at the pH value of 8.0, and the activity fluctuation of the cellulase reaches 80%.
Example 3 experiment on the washfastness after the fabric "refreshment" finish:
the experimental method comprises the following steps:
fabric washing pretreatment: two pieces of the same bright blue 30S carded yarn all-cotton knitted double-sided cloth (more hairiness on the cloth surface) are adopted, and edges are sewn on the periphery. One of the blocks is washed with 2g/L brightener at 30 ℃ for 20min, then dehydrated, dried and then ironed by an iron for flattening. And washing another piece of cloth with 2g/L standard washing powder containing 1.0% cellulase in a washing machine for 15 times continuously, washing with water at 30 deg.C for 30min each time, dewatering, oven drying, and ironing with an iron.
The washing process comprises the following steps: and (3) simultaneously putting the knitted double-sided cloth which is respectively treated by the brightening agent and treated by the enzymatic detergent and has the same appearance into a washing machine, washing the knitted double-sided cloth for 30min by adopting 1g/L standard detergent at 30 ℃, dehydrating, washing the knitted double-sided cloth twice at normal temperature for 5min each time, dehydrating and automatically drying the knitted double-sided cloth. Washing was repeated 20 times as described above. Comparing the cloth cover hairiness effects after washing for 5 times, 10 times, 15 times and 20 times respectively, wherein the cloth cover hairiness effects adopt a rating mode:
the results are shown in Table 3:
TABLE 3
| Number of washes | 0 | 5 | 10 | 15 | 20 |
| Washing treatment (rating) with the thermophilic chaetomium cellulase washing powder | 4-5 stages | 4-5 stages | 4-5 stages | 4-5 stages | 4 stage |
| Brightener treatment (rating) | 4-5 stages | Grade 3-4 | 2-3 stages | Stage 2 | 1-2 stages |
The cloth surface hairiness before the fabric is washed is determined as grade 1, the surface hairiness is completely removed is determined as grade 5 (the effect of returning new is best), and the specific grading conditions are as follows:
level 1: before treatment, the cloth surface has more dense filoplume
And 2, stage: the hair feather on the cloth surface is slightly removed, but a plurality of hair feathers and fuzzy lines still exist;
and 3, level: the hair feather on the cloth surface is obviously removed, but still has a certain amount of hair feather and a certain pattern;
4, level: most of the surface hairiness is removed, the surface has only a small amount of fluff, and the grains are clear;
and 5, stage: when the cloth cover hairiness is completely removed, the cloth cover hairiness is grade 5, and the surface has almost no hairiness;
2-3 level: the cloth cover hairiness is between 2-level and 3-level cloth cover hairiness;
as can be seen from the above experiments, after the fabric treated by the brightener is washed for 20 times, the fabric generates a plurality of hairiness, and the effect of 'renewing' is poor; after the fabric is washed and treated by standard washing powder containing 1.0 percent of the thermophilic chaetomium cellulase, the fabric has better refreshing effect after being washed for 20 times. The addition of Chaetomium thermophilum cellulase to the detergent results in a durable 'refreshment' treatment of the fabric.
Example 4 experiment on the "refreshment" finishing of the fabric:
the experimental method comprises the following steps:
eight pieces of the same bright blue 30S carded yarn all-cotton knitted double-sided cloth are taken, and edges are sewn on the periphery of the cloth. Then divided equally into two groups of four blocks each. One group was washed with 0.004% Chaetomium thermophilum cellulase, and the other group was washed with 3.000% of the commercially available cellulase YL400E (available from Guangzhou, Atlantic Biotech, Inc.). Washing with water at 30 deg.C under pH 8.5 for 360min, respectively taking out one block at 90min, 180min, 270min and 360min, dehydrating, and oven drying. The hair feather effect of the cloth surface adopts a rating mode:
the results are shown in Table 4:
TABLE 4
| Time of washing | 0 | 90min | 180min | 270min | 360min |
| Chaetomium thermophilum cellulase | 1 | 2－3 | 3－4 | 4－5 | 4－5 |
| Commercial cellulase YL400E | 1 | 2 | 3 | 4 | 4－5 |
From the above experiments, it can be seen that the removal effect of the thermophilic chaetomium cellulase on cotton fabric hairiness is better than that of the cotton fabric hairiness when the dosage of YL400E of the cellulase is 750 times at 30 ℃ and under the condition of pH value of 8.5. The thermophilic chaetomium cellulase is used in the detergent, the dosage of the thermophilic chaetomium cellulase is 1/750 of the cellulase YL400E, and the cost of the enzyme-added detergent can be greatly reduced.
Example 5:
1.0 percent of thermophilic chaetomium cellulase is added into standard washing powder, and the washing powder is used for washing brilliant blue 30S carded yarn all-cotton knitted double-sided cloth (more hair on the cloth surface) at the temperature of 30 ℃. The washing powder containing 1.0% of Chaetomium thermophilum cellulase is used in an amount of 1g/L in each washing, the pH value is 10.0-10.5 in each washing, the washing time is 30min, and then the washing powder is washed twice, dehydrated and dried, and washed for 15 times according to the method. The cloth surface hairiness effect reaches 4-5 grade from 1 grade.
Example 6:
1.0% thermophilic chaetomium cellulase is added into a standard laundry detergent, and the bright blue 30S carded yarn all-cotton knitted double-sided cloth (more hair feathers on the cloth surface) is washed by the laundry detergent at the temperature of 30 ℃. The usage amount of the thermophilic chaetomium cellulase laundry detergent containing 1.0% is 1g/L in each washing, the pH value is 8.0-8.2 in the washing, the washing time is 30min, and then the laundry detergent is washed twice by water, dehydrated and dried, and is washed for 10 times by the method. The cloth surface hairiness effect reaches 4-5 grade from 1 grade.
Example 7:
0.1 percent of thermophilic chaetomium cellulase is added into a standard laundry detergent, and the laundry detergent is used for washing brilliant blue 30S carded yarn all-cotton knitted double-sided cloth (more hair feathers on the cloth surface) at the temperature of 50 ℃. The dosage of the thermophilic chaetomium cellulase laundry detergent containing 0.1 percent is 1g/L during each washing, the pH value during the washing is 8.0-8.2, the washing time is 30min, and then the laundry detergent is washed twice by water, dehydrated and dried, and is washed 20 times by the method. The cloth surface hairiness effect reaches 4-5 grade from 1 grade.
Example 8:
2.0 percent of thermophilic chaetomium cellulase is added into washing powder, and the washing powder is used for washing brilliant blue 30S carded yarn all-cotton knitted double-sided cloth (more hair on the cloth surface) at the temperature of 15 ℃. The washing powder containing 2.0% of Chaetomium thermophilum cellulase is used in an amount of 1g/L in each washing, the pH value in the washing is 10.8-11.0, the washing time is 30min each time, and then the washing powder is washed twice, dehydrated and dried, and washed 10 times according to the method. The cloth surface hairiness effect reaches 4-5 grade from 1 grade.
Example 9:
0.3 percent of thermophilic chaetomium cellulase is added into washing powder, and the washing powder is used for washing brilliant blue 30S carded yarn all-cotton knitted double-sided cloth (more hair on the cloth surface) at the temperature of 20 ℃. The washing powder containing 2.0% of Chaetomium thermophilum cellulase is 3g/L in each washing, the pH value is 10.8-11.0 in each washing, the washing time is 30min, and then the washing powder is washed twice, dehydrated and dried, and is washed 10 times according to the method. The cloth surface hairiness effect reaches 4-5 grade from 1 grade.
Example 10:
0.3 percent of thermophilic chaetomium cellulase is added into a laundry detergent, and the washing detergent is used for washing brilliant blue 30S carded yarn all-cotton knitted double-sided cloth (more hair on the cloth surface) at the temperature of 40 ℃. The dosage of the thermophilic chaetomium cellulase washing powder containing 2.0 percent is 0.5g/L during each washing, the pH value during the washing is 10.8-11.0, the washing time is 30min, and then the washing is carried out twice, the dehydration and the drying, and the washing is carried out for 10 times according to the method. The cloth surface hairiness effect reaches 4-5 grade from 1 grade.
Sequence listing
<110> Hunan Eurter Biochemical Co., Ltd, Shanghai Eurter Biochemical Co., Ltd
<120> application of chaetomium thermophilum cellulase in preparation of detergent
<130>WH-NP-18-100019
<160>3
<170>Patent in version 3.4
<210>1
<211>1132
<212>DNA
<213> Chaetomium thermophilum
<400>1
atgcggtcga ctcctgttct ccgtaccgcc cttgcggctg ctctcccctt cactgtcctg 60
gctgccgacg gcaagtccac taggtactgg gactgctgca agccttcttg ctcgtggccc 120
ggcaaggctg ctgtgagcca acccgtcttc gcctgtgacc gcaacttcaa ccgcatctat 180
gacttcggtg ccaagtctgg ctgcgagggc ggtccggcct attcttgcgc cgaccagacc 240
ccgtgggctg tcaacgacca attctcgtac ggcttcgctg ccaccaacat tgccggcggt 300
aacgaggctt catggtgctg cgcttgctac aagtgagttc ttgcgtttcc aagaatagcc 360
aagggatctc ggctaatgca ctttaggctc accttcacct cggggcccgt ggccggcaag 420
gtcatggttg tccagtcgac cagcacgggc ggtgaccttg gcaacaacca tttcgacctg 480
aacatcccag gtggaggcgt tggtatcttc gatggttgca cgccccagtt cggcggtctg 540
cccggcgagc ggtacggcgg gatctcgtcg cgcagccagt gcgacagctt cccggatgcc 600
ctcaagcctg gctgctactg gcgcttcgac tggttcctga acgctgacaa cccgaacttc 660
accttcgagc gcgtccagtg tccttccgag cttgttgccc gcaccggctg caagcgcaat 720
gacgacggca actaccccgt cttcactcct ccttcgggag acagccccag cagcagcagc 780
gctgctccta cctccacctc gactgtaagt cttattctta cggaggccaa ggatacactt 840
ggatgatctc cttgctaacg ataattttcc tacagtcgca gcagccgcag cagccgacct 900
ccagcagctc gcaggcttct gtgccgacta gcaaccctgg tggctgcacc tctcaaaagt 960
gggctcagtg cggcggcatt ggcttcactg gctgcactac ctgcgtctcg ggcaccactt 1020
gcaccaagct gaatgactgg tactcgcagt ggtatgtctt atacggttct gtctctgggc 1080
gtgatcagca gctaacatta cgcagcttgt aagcacaatg atcaacctgt ga 1132
<210>2
<211>945
<212>DNA
<213> Artificial sequence (unknown)
<400>2
atgcggtcga ctcctgttct ccgtaccgcc cttgcggctg ctctcccctt cactgtcctg 60
gctgccgacg gcaagtccac taggtactgg gactgctgca agccttcttg ctcgtggccc 120
ggcaaggctg ctgtgagcca acccgtcttc gcctgtgacc gcaacttcaa ccgcatctat 180
gacttcggtg ccaagtctgg ctgcgagggc ggtccggcct attcttgcgc cgaccagacc 240
ccgtgggctg tcaacgacca attctcgtac ggcttcgctg ccaccaacat tgccggcggt 300
aacgaggctt catggtgctg cgcttgctac aagctcacct tcacctcggg gcccgtggcc 360
ggcaaggtca tggttgtcca gtcgaccagc acgggcggtg accttggcaa caaccatttc 420
gacctgaaca tcccaggtgg aggcgttggt atcttcgatg gttgcacgcc ccagttcggc 480
ggtctgcccg gcgagcggta cggcgggatc tcgtcgcgca gccagtgcga cagcttcccg 540
gatgccctca agcctggctg ctactggcgc ttcgactggt tcctgaacgc tgacaacccg 600
aacttcacct tcgagcgcgt ccagtgtcct tccgagcttg ttgcccgcac cggctgcaag 660
cgcaatgacg acggcaacta ccccgtcttc actcctcctt cgggagacag ccccagcagc 720
agcagcgctg ctcctacctc cacctcgact tcgcagcagc cgcagcagcc gacctccagc 780
agctcgcagg cttctgtgcc gactagcaac cctggtggct gcacctctca aaagtgggct 840
cagtgcggcg gcattggctt cactggctgc actacctgcg tctcgggcac cacttgcacc 900
aagctgaatg actggtactc gcagtgcaca atgatcaacc tgtga 945
<210>3
<211>314
<212>PRT
<213> Artificial sequence (unknown)
<400>3
MRSTPVLRTA LAAALPFTVL AADGKSTRYW DCCKPSCSWP GKAAVSQPVF ACDRNFNRIY 60
DFGAKSGCEG GPAYSCADQT PWAVNDQFSY GFAATNIAGG NEASWCCACY KLTFTSGPVA 120
GKVMVVQSTS TGGDLGNNHF DLNIPGGGVG IFDGCTPQFG GLPGERYGGI SSRSQCDSFP 180
DALKPGCYWR FDWFLNADNP NFTFERVQCP SELVARTGCK RNDDGNYPVF TPPSGDSPSS 240
SSAAPTSTST SQQPQQPTSS SSQASVPTSN PGGCTSQKWA QCGGIGFTGC TTCVSGTTCT 300
KLNDWYSQCT MINL 314
Claims (10)
1. The application of the chaetomium thermophilum cellulase in preparing the detergent, wherein the pH value of the chaetomium thermophilum cellulase is in the range of 8.0-11.0, and the detergent contains the chaetomium thermophilum cellulase; the thermophilic chaetomium cellulase is prepared by the following method:
1) synthesizing a chaetomium thermophilum cellulase gene, wherein the sequence of the synthesized chaetomium thermophilum cellulase gene is shown as SEQ ID NO.2, and the synthesized gene code is more in line with the gene code of the high-efficiency expression of the trichoderma reesei;
2) constructing thermophilic chaetomium cellulase expression plasmid;
3) transformation of trichoderma reesei and screening of thermophilic chaetomium cellulase high-yield strain to obtain thermophilic chaetomiumThe amino acid sequence of the cellulase is shown in SEQ ID NO.3, and the thermophilic chaetomium cellulase secreted outside the cells is the amino acid sequence from 22 th to 314 th sites of the sequence shown in SEQ ID NO. 3; the step 3) is specifically as follows: first, byEcoRIDegrading the plasmid constructed in the step 2) to form a plasmid fragment, and transferring the plasmid fragment into a Trichoderma reesei RUT C-30 host cell by adopting a chemical conversion method; then culturing on a PDA culture medium containing hygromycin B, inoculating colonies which grow vigorously on the PDA culture medium, transferring spores to a fresh PDA culture medium through growth and germination to separate and purify transformants which are stable in transformation, inoculating the spores of the transformants to a trichoderma cellulase induction culture solution, after culturing, sampling and centrifuging, and taking supernatant to perform SDS-PAGE electrophoretic analysis; the pH value of the trichoderma cellulase induction culture solution is 4.8, and lactose is used as an inducer.
2. The use according to claim 1, wherein the thermophilic chaetomium cellulase has good capability of degrading the surface hairiness of cotton fabrics at the pH value of 8.0-11.0, and has good effect of removing the surface hairiness of the cotton fabrics.
3. The use according to claim 1, wherein the thermophilic chaetomium cellulase has relatively small enzyme activity fluctuation within the pH value range of 8.0-11.0, the enzyme activity fluctuation is within 30%, and the removal effect on the surface hairiness of cotton fabrics is stable.
4. The use according to claim 1, wherein the Chaetomium thermophilum cellulase is present in the detergent in an amount in the range of 0.1-2.0% by mass.
5. The use according to claim 4, wherein the Chaetomium thermophilum cellulase is present in the detergent in an amount in the range of 0.3-1.0% by weight.
6. The use according to claim 1, wherein the thermophilic chaetomium cellulase is adapted to operate at a temperature of from 15 ℃ to 50 ℃ in the context of detergent use.
7. The use according to claim 6, wherein the thermophilic Chaetomium cellulase is adapted to operate at a temperature of 20-40 ℃ in the context of detergent use.
8. Use according to claim 1, wherein the detergent has the effect of improving the washfastness of the fabric after refreshing.
9. Use according to claim 1, wherein the detergent has a wash time of 30min to 600 min.
10. The use according to claim 1, wherein the detergent is used in an amount ranging from 0.5 to 3.0 g/L.
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