CN113755388B

CN113755388B - Enterobacter mori LRZ01 strain for degrading cellulose and application thereof

Info

Publication number: CN113755388B
Application number: CN202111162932.2A
Authority: CN
Inventors: 林文珍; 徐炜; 郭莺; 林志楷; 周平; 刘如振; 郭迟鸣; 孟红岩; 刘黎卿; 陈菲; 陈逍遥
Original assignee: SUBTROPICAL CROPS INSTITUTE OF FUJIAN PROVINCE
Current assignee: SUBTROPICAL CROPS INSTITUTE OF FUJIAN PROVINCE
Priority date: 2021-09-30
Filing date: 2021-09-30
Publication date: 2023-03-14
Anticipated expiration: 2041-09-30
Also published as: CN113755388A

Abstract

The invention discloses a strain of Enterobacter mori LRZ01 for degrading cellulose and application thereof. The Enterobacter mori LRZ01 is preserved in Guangdong province microorganism strain collection center at 2021, 9 and 2 days, and the preservation number is GDMCC No. 61916. The Enterobacter mori LRZ01 is derived from a mixture of pericarp and pulp of healthy chickpea, and has degradation capability on various biologically derived celluloses, such as sodium carboxymethylcellulose, rice straws, filter paper sheets, bamboos, tea stalks, barks and the like.

Description

Enterobacter mori LRZ01 strain for degrading cellulose and application thereof

Technical Field

The invention relates to the field of microbial strains, in particular to a strain of Enterobacter mori LRZ01 for degrading cellulose and application thereof.

Background

Cellulose is a macromolecular polysaccharide composed of glucose, is insoluble in water and common organic solvents, and is the main component of plant cell walls. Cellulose is a polysaccharide which is widely distributed and has the largest content in the nature, and accounts for more than 50 percent of the carbon content in the plant. The cellulose content of cotton is close to 100%, and is the purest cellulose source in nature. In general, the content of cellulose is 40-50%, and the content of hemicellulose is 10-30% and the content of lignin is 20-30%. Lignocellulose as the most abundant renewable resource on the earth is difficult to degrade, which not only causes resource waste, but also causes environmental pollution.

The current methods for treating cellulose mainly include chemical, physical and biological methods. The biological treatment method has mild conditions, belongs to an environment-friendly treatment method, and is a cellulose treatment method with great potential. The method for treating straws by adopting microbial technology is the most studied method for treating straws at present. Currently, most commonly studied are molds, typically Trichoderma viride, trichoderma reesei, and Trichoderma koningii [ Robson LM, chammass GH.Cellulases of bacterial origin [ J ]. Enzyme Microb Technol,1989,11 (10): 626-644;

lynd LR, weimer PJ, van Zyl WH, et al, microbial cell utilization, fundamentals and biotechnology [ J ] microbial Mol Biol Rev,2002,66 (3): 506-577 ], with less research on bacteria and actinomycetes. The domestic patents related to the bacteria capable of degrading cellulose mainly include a strain of lignocellulose substance efficient degrading bacteria L124 and application thereof related to Enterobacter hopcalis (Enterobacter hormaechei) separated from soil, and the application is 2016,3,9 and CN 105385628A; a strain of lignocellulose substance efficient degradation bacterium K24 and application thereof are disclosed in application publication 2016,1,27, application publication No. CN 105274037A; a new strain of Enterobacter, its preparation method and application are disclosed in publication No. 2017, 12,08, application publication No. CN 107446834A, which relates to Enterobacter sp obtained from rumen of cattle; escherichia coli (Escherichia coli) obtained from rumen of cattle is related to Escherichia coli (Escherichia coli) which can secrete cellulase and its application in degrading cellulose, and has publication No. CN 111733103A (2020, 10, 02).

While the patents related to Enterobacter mori (Enterobacter mori) mainly include Enterobacter mori and methods for producing natural vanillin by biotransformation of ferulic acid using the Enterobacter mori, application publication No. 2013,7,24, application publication No. CN 103215197A; enterobacter mori and a method and application thereof for biologically converting and fermenting bran extract for cigarettes, which are published as 2013,7,31 and CN 103224894A on the application publication date; a preparation method and application of aroma-producing bacteria fermented flos Lonicerae spice are disclosed in application publication No. 2016, no. 6, no. 8, no. CN 105647644A; enterobacter mori and application thereof in degrading thiamethoxam are disclosed in application publication Nos. 2019,8 and 27 and CN 110172430A. The above patent mainly uses Enterobacter mori (Enterobacter mori) in the technical field of producing perfume and degrading pesticide-thiamethoxam, and there is no patent and literature report related to degrading cellulose.

Disclosure of Invention

The invention aims to provide a strain of Enterobacter mori LRZ01 for degrading cellulose and application thereof.

In order to achieve the aim, the invention provides an Enterobacter mori LRZ01, wherein the Enterobacter mori LRZ01 is preserved in Guangdong province collection center of microorganism strains at 2021, 9 and 2 days, and the preservation number is GDMCC No. 61916.

The invention also protects the application of the Enterobacter mori LRZ 01.

Further, the Enterobacter mori LRZ01 has a use of degrading cellulose.

Further, the Enterobacter mori LRZ01 has a use of producing CMC enzyme.

Further, the Enterobacter mori LRZ01 has the use of producing FPA enzyme.

Further, the Enterobacter mori LRZ01 has the application of producing bamboo protocellulase.

Furthermore, the cellulose is derived from sodium carboxymethylcellulose, rice straws, filter paper sheets, bamboos, tea stems and barks.

The 16SrDNA sequence of Enterobacter mori (Enterobacter mori) LRZ01 was 1447bp in length, and BLAST at NCBI showed 99.02% similarity to the Enterobacter mori of NR.146667. The biological characteristics of Enterobacter mori LRZ01 are: the bacterial colony forms a transparent ring on a carboxymethyl fiber-Congo red culture medium plate, is a gram-negative bacterium, is in a milky round shape, has a wet and smooth surface, and is in a rod shape; physiological and biochemical characteristics: the strain has good growth state at about 37 ℃ and pH of about 7.0, strong adaptability, can resist NaCl 7%, has heat resistance of 45 ℃, is continuously subcultured for 15 times, and has stable growth condition, enzyme production condition and enzyme activity.

The Enterobacter mori LRZ01 is derived from a mixture of peels and pulps of healthy chickpeas, has degradation capability on various biologically derived celluloses such as sodium carboxymethylcellulose, rice straws, filter paper sheets, bamboos, tea stems, barks and the like, and shows that the strain is rich in multi-component enzyme systems and contains CMC enzyme, FPA enzyme, bamboo protocellulase and the like.

Drawings

FIG. 1 is a photograph showing the front and back sides of a colony of Enterobacter mori (Enterobacter mori) LRZ01 plate.

FIG. 2 is a photograph of a transparent circle formed by degrading cellulose when Enterobacter mori (Enterobacter mori) LRZ01 was cultured for 48 hours.

FIG. 3 is a photograph of a transparent circle formed by degrading cellulose by Enterobacter mori (Enterobacter mori) strain LRZ01 when cultured for 48 hours in a rescreening culture.

FIG. 4 is a photograph of a transparent circle formed by degrading cellulose by continuous culture of Enterobacter mori (Enterobacter mori) strain LRZ01 for 4 days.

FIG. 5 is a phylogenetic tree diagram based on the 16S rDNA sequence of Enterobacter mori (Enterobacter mori) strain LRZ 01.

FIG. 6 is a graph of a glucose standard.

FIG. 7 is a graph showing the effect of substrates of different culture times and different pH on the activity of LRZ01 CMC enzyme of Enterobacter mori (Enterobacter mori).

FIG. 8 is a graph showing the effect of different culture times on the activity of Enterobacter mori (Enterobacter mori) LRZ01 FPA enzyme.

FIG. 9 is a graph showing the effect of different culture times on the activity of Enterobacter mori (Enterobacter mori) LRZ01 bamboo fibril enzyme.

Detailed Description

Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative and intended to explain the present invention and should not be construed as limiting the present invention. Those skilled in the art will recognize that the specific techniques or conditions, not specified in the examples, are according to the techniques or conditions described in the literature of the art or according to the product specification. The reagents or instruments used are conventional products which are commercially available, and are not indicated by manufacturers.

The culture medium comprises the following components:

1. LB solid Medium (g/L): 10.0 parts of peptone, 5.0 parts of yeast extract powder, 10.0 parts of NaCl, 15.0 parts of agar powder and natural pH;

2. selection medium (g/L) CMC-Na 5.0, mgSO4 0.3, K2HPO4 1.0, naH2PO4 1.0, agar 20.0, congo red 0.2, caCl2.2H2O 0.3, feSO4.7H2O 0.005, mnSO4.001, znC12.001, coC12.001, natural pH;

3. enzyme activity culture medium (g/L) including CMC-Na 5.0, peptone 2.5, yeast extract 0.5, mgSO ₄ 0.3,KH ₂ PO ₄ 2.0,NaCl 1.0,(NH4) ₂ SO ₄ 1.4,CaCl ₂ ·2H ₂ O 0.3,FeSO ₄ ·7H ₂ O 0.005,MnSO ₄ 0.001 6,ZnC1 ₂ 0.001 7,CoC1 ₂ 0.001 7, natural pH;

4. cellulose spectrum culture medium (g/L) carbon source (bamboo, straw, na) ₂ CO ₃ )5.0,MgSO ₄ 0.3,KH ₂ PO ₄ 2.0,NaCl 1.0,(NH4) ₂ SO ₄ 1.4,CaCl ₂ ·2H ₂ O 0.3,FeSO ₄ ·7H ₂ O 0.005,MnSO ₄ 0.001 6,ZnC1 ₂ 0.001 7,CoC1 ₂ 0.001 7, natural pH;

5. preparing different carbon sources, namely selecting cellulose acetate, filter paper, tea stalks, rice straws, fresh and tender bamboo stems and barks as the different carbon sources. Respectively cutting rice straw and bamboo into 2-3 cm small sections, drying at 90 deg.C, and pulverizing to 40 mesh for use.

The culture medium is sterilized at 121 deg.C for 20min.

Example 1: isolation and purification of Enterobacter mori (Enterobacter mori) LRZ01

Selecting healthy chickpeas, grinding the peels and pulps in a clean bench by using a mortar, transferring into a 500mL shaking flask, adding 200mL sterile water, and performing shaking culture at 28 ℃ and 160r/min for 24h. The supernatant was aspirated and diluted to 10 degrees in a gradient ^-1 、10 ^-2 、10 ^-3 、10 ^-4 、10 ^-5 、10 ^-6 And spreading the diluted solution with each concentration on a 9cm screening culture medium plate, and culturing for 12-48h at the constant temperature of 28 ℃ in a dark place. During the culture period, selecting the bacterial strain with obvious degradation ring around the colony from the screening culture medium, and performing three-stage streak inoculation and purification on an LB culture medium plate until a single colony is obtained.

See fig. 1. FIG. 1 is a front and back view of a colony of Enterobacter mori (Enterobacter mori) LRZ01 plate.

Example 2 screening of Enterobacter mori (Enterobacter mori) LRZ01

Inoculating the separated and purified strain into a plate of a screening culture medium, and culturing for 2-4 days in a constant-temperature incubator at 28 ℃ in a dark place. After 48h, the culture dish (9 cm) is observed, strains with transparent circles around the colonies are selected and marked, and the diameter D of the transparent circles and the diameter D of the colonies are measured at the same time. Finally, the strain LRZ01 (see fig. 2) with the largest transparent degradation circle (transparent circle diameter D-colony diameter D = degradability) was selected as the strain to be developed, and the strain was subcultured 15 times and inoculated again into a plate of the screening medium. After 48h, the diameter D of the clearing circle and the diameter D of the colony in the dish were measured (see FIG. 3). The diameter D of the transparent ring and the diameter D of the bacterial colony of the two experiments are not obviously changed, which shows that the growth condition, the enzyme production condition and the enzyme activity of the bacterial strain are stable and have no degradation phenomenon. FIG. 2 shows a strain map of Enterobacter mori LRZ01 before screening. FIG. 2 is a photograph of a transparent circle formed by degrading cellulose when Enterobacter mori (Enterobacter mori) LRZ01 was cultured for 48 hours. FIG. 3 is a diagram showing the strain of Enterobacter mori (Enterobacter mori) LRZ01 after selection. FIG. 4 is a photograph of a clearing circle formed by continuous culture of Enterobacter mori (Enterobacter mori) strain LRZ01 for 4 days to degrade cellulose (a photograph of a strain in which 3 colonies are inoculated at equal intervals on a 9cm dish for continuous culture for 4 days until the clearing circles formed by the 3 colonies overlap). A represents 1 day of culture, B represents 2 days of culture, C represents 3 days of culture, and D represents 4 days of culture.

Example 3 identification of Enterobacter mori LRZ01

(1) Morphological characteristics of the cells

Morphological identification of the strain LRZ 01: the purified strain LRZ01 is inoculated to an LB solid medium plate, and is cultured in an incubator at 37 ℃ in a dark place for 2 days. The results show that the bacterial colony of the strain LRZ01 is round, has a wet, smooth, milky white and opaque surface, irregular edge, uniform texture and flat shape.

(2) Molecular biological identification of 16S rNDA

Molecular biological identification of strain LRZ 01: and (3) performing small-scale fermentation and centrifugation to collect thalli of the strain LRZ01, washing the thalli with sterile water, adding a lysate, performing water bath for 10min, performing centrifugation at 10000rpm, and taking a supernatant to obtain the genomic DNA of the strain LRZ 01. The strain LRZ01 DNA was used as a template, and amplification was carried out according to the following reaction system and amplification conditions. The primer sequences are shown as SEQ ID NO 1 and SEQ ID NO 2, and the reaction system and the amplification conditions are respectively as follows:

the upstream primer 27F:5'-AGA GTT TGA TCC TGG CTC AG-3' SEQ ID NO;

the downstream primer 1492R:5'-TAC GGC TAC CTT GTT ACG ACT T-3' SEQ ID NO.

PCR reaction (20 uL): 2x PCR SuperMIx 10. Mu.L; 1 μ L of LRZ01 DNA template; 1 mu L of each of the upstream and downstream primers; ddH ₂ O 7μL。

PCR reaction procedure: pre-denaturation at 94 deg.C for 10min; denaturation at 94 deg.C for 1min; annealing at 55 deg.C for 1min; extending at 72 ℃ for 1min; circulating for 35 times; extending at 72 ℃ for 10min; keeping the temperature at 10 ℃.

The PCR products were detected by electrophoresis on 1.5% agarose and sequenced by the Biotech company, inc., of Shanghai Biotech, inc. The sequencing results were:

16S rDNA sequencing result of strain LRZ 01:

ACTTTGCGGCGGGCCTACCATGCAAGTCGAGCGGCAGCGGAAAGTAGCTTGCTACTTTGCCGGCGAGCGGCGGACGGGTGAGTAATGTCTGGGAAACTGCCTGATGGAGGGGGATAACTACTGGAAACGGTATCTAATACCGCATAACGTCGCAAGACCAAAGAGGGGGACCTTCGGGCCTCTTGCCATCAGATGTGCCCAGATGGGATTATCTAGTAGGTGGGGTAACGGCTCACCTAGGCGACGATCCCTAGCTGGTCTGAGAGGATGACCACCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCGCGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGCGGTGAGGTTAATAACCTCATCGATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTCTGTCAAGTCGGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATTCGAAACTGGCAGGCTAGAGTCTTGTAGAGGGGGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTGGACAAAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGTGGCTTCCGGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTACTCTTGACATCCAGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTCTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTGCCAGCGGTCCGGCCGGGAACTCAAAGGAGACTGCCAGTGATAAACTGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGAGTAGGGCTACACACGTGCTACAATGGCGCATACAAAGAGAAGCGACCTCGCGAGAGCAAGCGGACCTCATAAAGTGCGTCGTAGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTAGATCAGAATGCTACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCAAAAGAAGTAGGTAGCTTAACCTTCGGGAGGGCGCTACCACTTTGATTCAGGGGG。SEQ ID NO:3。

the obtained sequencing results were analyzed on-line by NCBI BLAST (https:// blast.ncbi. Nlm. Nih. Gov/BLAST. Cgi) and showed 99% sequence homology with the published related Enterobacter mori (Enterobacter mori) strain. And a phylogenetic tree (figure 5) is constructed by adopting software MEGA-X, and the strain LRZ01 has the closest evolutionary distance with Enterobacter (NR.146667).

The strain LRZ01 was deposited with the following deposit information:

the strain name: enterobacter mori LRZ01;

the preservation organization: guangdong province microbial strain preservation center;

and (4) storage address: building No. 59, building No. 5 of prefecture Zhonglu No. 100 yard, guangzhou city;

preservation time: 9/2/2021;

the preservation number is as follows: GDMCC No. 61916.

Example 4 utilization of different carbon sources by Enterobacter mori (Enterobacter mori) LRZ01

The CMC-Na in the screening culture medium is respectively changed into cellulose acetate, rice straws, filter paper sheets, bamboos, barks and tea stalks with equal mass, culture mediums with different carbon sources are prepared (preparation of different carbon sources, namely selecting the cellulose acetate, the rice straws, the filter paper sheets, the bamboos, the barks and the tea stalks as different carbon sources, the rice straws, the filter paper sheets, the bamboos, the barks and the tea stalks are respectively cut into 2cm-3cm sections, dried at 90 ℃ and then crushed into 40 meshes for later use), and the cellulose degrading strain LRZ01 obtained by separation and purification is cultured for 4 days at 3 points of each flat plate at 28 ℃. Staining the strain with 0.02% Congo red for 10min, washing with 1mol/L NaCl, measuring the diameter (D) of a transparent ring and the diameter (D) of a bacterial colony by using a vernier caliper, respectively, representing a degradation ring by using the difference value of the diameter of the transparent ring and the diameter of the bacterial colony, and preliminarily judging the degradation capability of the strain on different cellulose materials according to the size of the degradation ring.

The result shows that the strain LRZ01 has the degradation capability on CMC-Na, cellulose acetate, rice straws, filter paper sheets, bamboos, tree parts and tea stems, but has the strongest degradation capability on CMC-Na. The degradation capability of the strain LRZ01 on cellulose acetate is second to CMC-Na, the strain has stronger degradation capability on xylem, certain degradation capability on rice straw and bamboo, and weak degradation capability on filter paper and tea stalks, and the results are shown in Table 1.

TABLE 1 degradation ability of LRZ01 strain on different carbon sources

Note that the diameter of the transparent circle-the diameter of the colony = the degradation ability, the degradation ability is indicated by "+", and "-" indicates that the degradation circle is less than 5mm or has no degradation ability; the degradation ring is 35mm-40mm; the degradation ring is 30mm-35mm; the degradation circle is 25mm-30mm; the degrading ring is 20mm-25mm; the degradation circle is 15mm-20mm; the degradation circle is 10mm-15mm; the degradation ring is 5mm-10mm.

Example 5 determination of enzyme Activity by DNS method-drawing of glucose Standard Curve

Accurately weighing 104.8mg of glucose with constant weight, dissolving with distilled water to constant volume in a 100mL volumetric flask, and preparing into 1mg/mL standard glucose solution as standard glucose mother liquor for later use. The addition and reaction of each reagent was completed as in Table 2, and after cooling, the color was compared at 540nm, and the absorbance was recorded to draw a glucose standard curve (see FIG. 6).

TABLE 2 ingredient Table of glucose solutions of different concentrations

Definition of enzyme Activity the enzyme amount for hydrolyzing a substrate to generate 1. Mu.g of glucose per minute by 1mL of enzyme solution at 50 ℃ is referred to as 1 enzyme activity unit and is expressed in U/mL.

The method for calculating the enzyme activity comprises the following steps:

1 unit of enzyme activity X = 1000 xcxv/(V1 xt).

Wherein X is the enzyme activity (U/mL) of the sample; c, the content of glucose in the test solution (mg/10 mL); v is constant volume (mL); v1, the volume (mL) of the enzyme solution is aspirated; t is hydrolysis time (min).

Example 6 determination of Enterobacter mori (Enterobacter mori) LRZ01 CMC enzyme Activity

6.0mL of 1.0% sodium carboxymethyl cellulose solution (pH 7.0, 6.0 and 8.0) is added into a 10.0mL test tube, 2.0mL of crude enzyme solution with different culture times is added, enzymolysis is carried out for 30min in water bath at 50 ℃, 2.0mL of DNS solution is added, boiling water bath is carried out for 8min, the mixture is taken out and cooled to room temperature in ice water, the volume is determined to be 10.0mL, the absorbance is measured at 540nm, and the enzyme activity is calculated according to a glucose standard curve. The results are shown in FIG. 7.

FIG. 7 is a graph showing the effect of culture time and various pH substrates on CMC-producing enzymes of Enterobacter mori (Enterobacter mori) strain LRZ 01. The results of FIG. 7 show that the CMC enzyme produced by the strain LRZ01 has stronger enzyme activity under the reaction condition that the pH of a substrate is 8.0, the enzyme activity of the 7 th enzyme reaches the highest 403.70U/mL, and the enzyme activity starts to have a descending trend after 7 days.

Example 7 measurement of Enterobacter mori (Enterobacter mori) LRZ01 FPA enzyme Activity

Adding 1cm × 6cm filter paper into a 10.0mL test tube, adding 6.0mL of pH 7.0 buffer solution as a substrate, adding 2.0mL of crude enzyme solution with different culture times, performing enzymolysis at 50 ℃ for 30min, adding 2.0mL of DNS solution, performing boiling water bath for 8min, taking out, cooling to room temperature in ice water, diluting to 10.0mL, measuring the absorbance at 540nm, and calculating the enzyme activity according to a glucose standard curve. The results are shown in FIG. 8.

FIG. 8 is a graph showing the effect of culture time on FPA-producing enzyme of Enterobacter mori LRZ01 strain. The results in FIG. 8 show that the optimum culture time for the strain LRZ01 to produce FPA enzyme is 7 days, and the 7 th enzyme activity reaches the maximum value of 190.16U/mL and then decreases with the increase of fermentation days.

Example 8 measurement of Enterobacter mori (Enterobacter mori) LRZ01 bamboo Cellulosidase Activity

Inoculating strain LRZ01 to 50mL fermentation medium containing bamboo fibril as sole carbon source, performing shake culture at 28 deg.C for 1-10 days, and calculating enzyme degradation rate by difference weight method with no strain as blank control.

Enzymatic degradation rate (%) = [ (mass before enzymatic hydrolysis-mass after enzymatic hydrolysis)/mass before enzymatic hydrolysis ] × 100%.

The results are shown in FIG. 9, which is a graph showing the effect of the culture time of Enterobacter mori (Enterobacter mori) strain LRZ01 on the degradation rate of bamboo fibril.

As can be seen from fig. 9, the degradation ability of strain LRZ01 to bamboo fibril was enhanced with the increase of the culture time, strain LRZ01 tended to be stable from 8d to 9d, and the degradation rate reached the maximum of 20.8% at 9 d.

Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made in the above embodiments by those of ordinary skill in the art without departing from the principle and spirit of the present invention.

SEQUENCE LISTING

<110> research institute for subtropical plants in Fujian province

<120> Enterobacter mori LRZ01 strain for degrading cellulose and application thereof

<130> YRDZ-21001-CNI

<160> 3

<170> PatentIn version 3.5

<210> 1

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<213> Artificial sequence

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agagtttgat cctggctcag 20

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tacggctacc ttgttacgac tt 22

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actttgcggc gggcctacca tgcaagtcga gcggcagcgg aaagtagctt gctactttgc 60

cggcgagcgg cggacgggtg agtaatgtct gggaaactgc ctgatggagg gggataacta 120

ctggaaacgg tatctaatac cgcataacgt cgcaagacca aagaggggga ccttcgggcc 180

tcttgccatc agatgtgccc agatgggatt atctagtagg tggggtaacg gctcacctag 240

gcgacgatcc ctagctggtc tgagaggatg accacccaca ctggaactga gacacggtcc 300

agactcctac gggaggcagc agtggggaat attgcacaat gggcgcaagc ctgatgcagc 360

catgccgcgt gtatgaagaa ggccttcggg ttgtaaagta ctttcagcgg ggaggaaggc 420

ggtgaggtta ataacctcat cgattgacgt tacccgcaga agaagcaccg gctaactccg 480

tgccagcagc cgcggtaata cggagggtgc aagcgttaat cggaattact gggcgtaaag 540

cgcacgcagg cggtctgtca agtcggatgt gaaatccccg ggctcaacct gggaactgca 600

ttcgaaactg gcaggctaga gtcttgtaga ggggggtaga attccaggtg tagcggtgaa 660

atgcgtagag atctggagga ataccggtgg cgaaggcggc cccctggaca aagactgacg 720

ctcaggtgcg aaagcgtggg gagcaaacag gattagatac cctggtagtc cacgccgtaa 780

acgatgtcga cttggaggtt gtgcccttga ggcgtggctt ccggagctaa cgcgttaagt 840

cgaccgcctg gggagtacgg ccgcaaggtt aaaactcaaa tgaattgacg ggggcccgca 900

caagcggtgg agcatgtggt ttaattcgat gcaacgcgaa gaaccttacc tactcttgac 960

atccagagaa ctttccagag atggattggt gccttcggga actctgagac aggtgctgca 1020

tggctgtcgt cagctcgtgt tgtgaaatgt tgggttaagt cccgcaacga gcgcaaccct 1080

tatcctttgt tgccagcggt ccggccggga actcaaagga gactgccagt gataaactgg 1140

aggaaggtgg ggatgacgtc aagtcatcat ggcccttacg agtagggcta cacacgtgct 1200

acaatggcgc atacaaagag aagcgacctc gcgagagcaa gcggacctca taaagtgcgt 1260

cgtagtccgg attggagtct gcaactcgac tccatgaagt cggaatcgct agtaatcgta 1320

gatcagaatg ctacggtgaa tacgttcccg ggccttgtac acaccgcccg tcacaccatg 1380

ggagtgggtt gcaaaagaag taggtagctt aaccttcggg agggcgctac cactttgatt 1440

caggggg 1447

Claims

1. Enterobacter moriEnterobacter mori LRZ01, characterized in that said Enterobacter moriEnterobacter mori LRZ01 was deposited at the Guangdong province Collection of microorganisms at 2021, 9, 2 days, with the accession number of GDMCC No. 61916.

2. Enterobacter mori of claim 1Enterobacter mori Use of LRZ01 in degrading cellulose.

3. Use according to claim 2, wherein the cellulose is derived from sodium carboxymethylcellulose, rice straw, filter paper sheets, bamboo, tea stems, bark.

4. Enterobacter mori of claim 1Enterobacter mori Use of LRZ01 in CMC producing enzymes.

5. Enterobacter mori of claim 1Enterobacter mori Use of LRZ01 in the production of FPA enzyme.

6. Enterobacter mori of claim 1Enterobacter mori Use of LRZ01 in bamboo protocellulase production.