CN109161565A - A method of ethyl alcohol is produced using whey - Google Patents
A method of ethyl alcohol is produced using whey Download PDFInfo
- Publication number
- CN109161565A CN109161565A CN201811220905.4A CN201811220905A CN109161565A CN 109161565 A CN109161565 A CN 109161565A CN 201811220905 A CN201811220905 A CN 201811220905A CN 109161565 A CN109161565 A CN 109161565A
- Authority
- CN
- China
- Prior art keywords
- whey
- lactose
- saccharomyces cerevisiae
- gene
- glucose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention provides a kind of efficient method for utilizing whey production ethyl alcohol, and the method is by utilizing type saccharomyces cerevisiae engineered yeast stationary culture fermentation next life producing and ethanol in the culture medium containing whey or lactose using the whey that one plant of deposit number is CGMCC No.11223.S. cervisiae of the present invention, the galactose metabolism regulation of S. cervisiae itself is eliminated while alleviating glucose repression phenomenon, the bacterium can grow in the culture medium that whey concn is 120g/L (lactose content 53.1g/L), ferment producing and ethanol, fermentation period is 54h, Lactose in Whey utilization rate is 98.7%, and dehydrated alcohol is 49.7% (be equivalent to theoretical yield 92.3%) to lactose yield；It alleviates glucose simultaneously to check galactolipin, makes glucose and galactolipin while being utilized.
Description
The application is the divisional application of following patent application, the application number of original application: 2015106320883, the applying date:
2015-09-29, denomination of invention: plant height effect utilizes the saccharomyces cerevisiae engineered yeast and its construction method of whey producing and ethanol.
Technical field
The invention belongs to technical field of bioengineering, are related to the breeding of industrial microorganism, especially with one plant of whey benefit
The method for efficiently utilizing whey production ethyl alcohol with type saccharomyces cerevisiae engineered yeast.
Background technique
Whey refers to when manufacturing cheese or casein, isolates remaining extremely thin liquid after flocculate, is industry
The byproduct of cheese and casein is produced, the cheese of every production 1t generates 9t whey, containing in milk 55% nutrition, but by
There is very high BOD and COD in whey, so causing very big burden to environment.Currently, global whey yield is about 1,600,000,000
Ton, and only therein 50% is handled, and food, feed etc. are used for, remaining about 800,000,000 tons do not obtain effective benefit
With being excreted in nature, not only cause the pollution of environment, and also cause the huge waste of the resource.In whey in addition to
Outside containing 20% milk protein, content it is most be lactose, account for about whey 5% or so, be the main original for generating high BOD and COD
Cause.
How using the lactose in whey, so that lowering whey is always asking for Dairy Industry concern to the pollution of environment
Topic.Utilizing mainly for lactose has following direction: for food service industry, being used for feedstuff industry, for pharmaceuticals industry.But with
The aggravation and the factors such as world food safety problem of global energy crisis, whey become the master of new production alcohol fuel
Raw material is wanted, but also becoming a completely new hot spot using whey production alcohol fuel.
Existing more than the 40 years history of alcohol fuel of overseas utilization whey production at present, used strain mainly have Crewe dimension
Yeast and saccharomyces cerevisiae.Although kluyveromyces can utilize whey, the category yeast ethanol production is low, and resistance is poor, bacterium
Body increment is big, is not suitable for producing ethyl alcohol.Although saccharomyces cerevisiae is the best first choice of production ethyl alcohol, but wild type Saccharomyces ferment
Mother can not be that sole carbon source allows to fermented whey producing and ethanol, be always so how saccharomyces cerevisiae is transformed using whey
The research hotspot of scientific research personnel.External solution is broadly divided into following several at present: one, being to utilize protoplast fusion skill
Art merges saccharomyces cerevisiae and kluyveromyces, and screening not only can use lactose but also the mutant strain of the efficient producing and ethanol of energy；
It two, is foreign gene to be imported into saccharomyces cerevisiae body using genetic engineering means, so that saccharomyces cerevisiae has and to reduce lactose
Ability, to solve the problems, such as that it cannot utilize whey.The LAC4 base that most foreign genes is kluyveromyces is imported at present
Because of (coding whey catabolic enzyme) and LAC12 gene (coding whey permease).But bacterial strain constructed by above two method is equal
There are wheys to utilize the problems such as slow, fermentation period is long.
For Lac+For saccharomyces cerevisiae engineered yeast strain, the lactose in whey to be first broken down into glucose and galactolipin it
It can just be further continued for being metabolized afterwards.And for saccharomyces cerevisiae, there are greatest differences when using galactolipin and glucose, this
Difference had both included the regulation of galactose metabolism itself, also included inhibition of the glucose to galactose utilization enzyme system, i.e. glucose hinders
Hold back phenomenon." substitution carbon source " of the galactolipin as saccharomyces cerevisiae, its metabolic rate is only glucose metabolism rate under oxygen consumption state
1/3.Galactolipin need to first pass through Leloir approach and be metabolized to just can enter glycolytic pathway after glucose 6-phosphate by yeast benefit
With.Enzyme needed for Leloir approach is encoded by GAL gene, and the GAL gene of saccharomyces cerevisiae belongs to typical Expression modulation gene.Have four
Kind gene product takes part in this Expression modulation of GAL genic system, they are the GAL4 gene on XVI chromosome respectively
Encode a kind of Gal4 albumen that can combine the upstream sequence site of above-mentioned five kinds of genes and activate these genetic transcriptions；Also it is located at
The Gal80 albumen of GAL80 gene coding on chromosome x III can then be bound directly with Gal4 albumen, to inhibit turning for Gal4
Record activation function；And the Gal3 albumen of the GAL3 gene coding on chromosome IV can be in the work of galactolipin inducement signal
Allosteric effect occurs with lower same ATP, galactolipin, the allosteric effector of generation discharges Gal4 albumen with Gal80 protein binding, but
Be current galactolipin generate inducement signal mechanism of action it is not clear.Other than above-mentioned three kinds of modulins, table is studied recently
The Gal6 albumen of the bright GAL6 gene coding on chromosome x IV also belongs to one kind of GAL controlling gene, and Gal6 albumen can shadow
Ring the stability of GAL family gene transcript mRNA.
In addition to GAL gene oneself expression adjustment process, expression is also inhibited by glucose.In saccharomycete, many bases
The expression of cause is by the control and adjustment effect of glucose, and checking for glucose is related to several factors, including glucose signals pass
Defeated, the intermediate regulations factor, special transcription repression and activator protein etc., Mig1 protein complexes and GAL- activator protein (Gal4) it
Between complexing action be considered as glucose repression galactose metabolism key step.
In saccharomyces cerevisiae, neutral seaweed carbohydrate-splitting enzyme is encoded by NTH1 and NTH2 gene, wherein playing a major role
It is the albumen of NTH1 coding.And the compound that glucose repression albumen composition is made of three albumen, wherein only by
The Mig1 albumen of MIG1 coding is combined with enzyme gene needed for galactose utilization, inhibits its transcription.And regulate and control way in galactose metabolism
In diameter, play negative regulation expressional function modulin altogether there are two, respectively Gal80 (being encoded by GAL80 gene) and Gal6
(being encoded by GAL6 gene).
Zou waits " Lac quietly+The influence that GAL80 gene knockout utilizes whey in saccharomyces cerevisiae engineered yeast " disclose building one
Strain can efficiently utilize the saccharomyces cerevisiae engineered yeast strain of whey, clone the GAL80 gene of AY5, and building knocks out plasmid pUC-
GABCUP.Using the plasmid as template PCR amplifications homologous recombination segment GA-CUPl-GB, with Lac+Saccharomyces cerevisiae AY51024M be by
Body bacterial strain, using homologous recombination method knock out F-strain in GAL80 gene, obtain one plant not only released Gal80 inhibit but also
The strains A Y51024M-G of whey can be utilized.
Currently, releasing the report of galactose metabolism regulation again simultaneously in overexpression LAC4 and LAC12 gene not yet both at home and abroad
Road.
Summary of the invention
The purpose of the present invention is efficiently utilize cream by providing one kind using type saccharomyces cerevisiae engineered yeast using one plant of whey
The method of clear production ethyl alcohol.
The method is to contain whey or lactose using the Wine brewing yeast strain that deposit number is CGMCC No.11223
Culture medium in stationary culture ferment next life producing and ethanol, fermentation period 54h.
Preferably, the composition of the culture medium are as follows: whey powder 120g/L or lactose 53.1g/L, (NH4)2SO45g/L,
MgSO4·7H2O 1g/L, solvent are water.
Preferably, the temperature of the fermentation is 30 DEG C.
Whey provided by the invention using type saccharomyces cerevisiae engineered yeast (Saccharomyces cerevisiae) be specially
AY5MG has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (referred to as on August 11st, 2015
CGMCC, address are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101),
Deposit number is CGMCC No.11223, classification naming: Saccharomyces Cerevisiae in S accharomyces cerevisiae.
Saccharomyces cerevisiae engineered yeast AY5MG of the present invention is 120g/L (lactose content 53.1g/L) in whey concn
Growth fermentation producing and ethanol in culture medium, fermentation period 54h, Lactose in Whey utilization rate are 98.7%；Dehydrated alcohol is to lactose
Yield is 49.7% (be equivalent to theoretical yield 92.3%).
Whey of the present invention utilizes the construction method of type saccharomyces cerevisiae engineered yeast, is by strong promoter PGK1 difference table
Up to galantase gene LAC4 and lactose permease gene LAC12, while MIG1, NTH1 and GAL6 gene are knocked out, slow
The galactose metabolism regulation of S. cervisiae itself is eliminated while solving glucose repression phenomenon, obtaining one plant has high patience
Efficiently utilize lactose producing and ethanol saccharomyces cerevisiae engineered yeast.
The construction method of saccharomyces cerevisiae engineered yeast strain of the present invention specifically comprises the following steps:
1) using saccharomyces cerevisiae genome as template, by PCR amplification go out GAL6 gene upstream and downstream segment GAL6A and
GAL6B connect to obtain plasmid pUC-G6AB with pUC19 plasmid；
2) it will be connect from the copper resistant gene C UP1 of Yep-C plasmid with pUC-G6AB, obtain plasmid pUC-G6AB-
CUP1；
3) homologous recombination of GAL6 gene is amplified using plasmid pUC-G6AB-CUP1 as template using PCR amplification
Segment GAL6A-CUP1-GAL6B；
4) GAL6A-CUP1-GAL6B homologous recombination segment is imported by saccharomyces cerevisiae recipient bacterium using lithium acetate transformation method
The a type of strain and α type monoploid, the genes of brewing yeast engineering haploid strains after obtaining homologous recombination；The F-strain is
Whey is expressed by strong promoter PGK1 using method disclosed in patent CN102604849B respectively and decomposes enzyme gene LAC4 and cream
Clear permease gene LAC12, while knocking out MIG1 gene and NTH1 gene and the genetic engineering bacterium that obtains.
5) by after purification saccharomyces cerevisiae a type and α type recombination monoploid merge, pass through resistant panel and the experiment of raw spore
Screening obtains can use the saccharomyces cerevisiae engineered yeast (amphiploid) of whey.
The present invention also provides a kind of dedicated for identifying the verifying sequence for removing galactose metabolism negative regulator,
The gene order is to utilize Wine-making Using Whey Yeast engineering bacterium strain genome as mould using described using G6A-U and G6B-D as primer
Plate, amplified fragments sequencing is a specific sequence, as shown in sequence table 1.
The primer size that G6A-U and G6B-D are amplified be can use to judge whether the gene is removed.
The advantages and positive effects of the present invention:
The present invention makes bacterial strain on the basis of original utilization whey ability, holds back member by blocking galactose metabolism to regulate and control negative resistance
Part gene, the saccharomyces cerevisiae engineered yeast strain for obtaining one plant of removal galactose metabolism regulation, rapidly and efficiently utilizing whey producing and ethanol
AY5MG。
The present invention saccharomyces cerevisiae engineered yeast Saccharomyces cerevisiae obtained that can use whey
AY5MG (deposit number CGMCC No.11223) and initial S. cervisiae (recipient bacterium AY-510B24M Saccharomyces
Cerevisiae CGMCC No.5843) it compares, whey more can be quickly utilized, is 120g/L (lactose in whey concn
Content is 53.1g/L) culture medium in growth, fermentation producing and ethanol；Fermentation period is 54h, and Lactose in Whey utilization rate is
98.7%；Dehydrated alcohol is 49.7% (be equivalent to theoretical yield 92.3%) to lactose yield；Saccharomyces cerevisiae is alleviated simultaneously
The galactose metabolism regulation of itself, utilizes whey quickly, is being containing 19% (v/v) ethyl alcohol or fermentation temperature
It can be with normal fermentation at 39 DEG C.Saccharomyces cerevisiae engineered yeast of the present invention does not have particular/special requirement, general brewery to Zymolysis Equipment and condition
Equipment and condition can be used, thus have wide practical use, can provide for the production alcohol fuel by raw material of whey can
Energy.
Detailed description of the invention
Fig. 1: plasmid pUC-G6AB-CUP1 building process；
Fig. 2: plasmid pUC-G6AB-CUP1 verifying electrophoretogram.
Fig. 3: the haploid verifying of positive restructuring saccharomyces cerevisiae.
Fig. 4: whey utilizes the building route map of type saccharomyces cerevisiae engineered yeast.
Fig. 5: engineering bacteria ferments with parent in simulation whey decomposed solution compared with situation, wherein (A) is parent AY-
510B24M, (B) are engineering bacteria AY5MG.
Specific embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
Under the premise of invention spirit and scope, to the various changes or change of material component and dosage progress in these embodiments
It belongs to the scope of protection of the present invention.
F-strain used in the present invention is the Lac that can use any source+Saccharomyces cerevisiae amphiploid bacterial strain.
Embodiment 1: the building of the Wine-making Using Whey yeast gene engineering bacteria of removal galactose metabolism regulation
(1) building of engineering strain
1) using saccharomyces cerevisiae AY5 (Saccharomyces cerevisiae CGMCC No 2.1364) genome as mould
Plate is the piece of 646bp size using the length that PCR amplification goes out GAL6 gene complete sequence upstream using G6A-U and G6A-D as primer
Section GAL6A is similarly primer using G6B-U and GB6-D, and the length for amplifying GAL6 gene complete sequence downstream is 665bp size
Segment GAL6B (primer sequence and restriction enzyme site are shown in Table 1), then connect GAL6A, GAL6B and pUC19 plasmid to obtain plasmid
pUC-G6AB；
2) using Yep-C plasmid as template, using Cup-U and Cup-D1, Cup-D2 as primer, the copper resistant of 1410bp is amplified
Gene C UP1 is connected with plasmid pUC-G6AB, obtains plasmid pUC-G6AB-CUP1 (building process is shown in Fig. 1)；
3) using the pUC-G6AB-CUP1 plasmid constructed as template, using G6A-U, G6B-D as primer, PCR amplification skill is utilized
Art amplifies homologous recombination segment GAL6A-CUP1-GAL6B；
Primer sequence and restriction enzyme site used in the process of 1 plasmid construction of table
Fig. 2 is the electrophoresis proof diagram of the building process of pUC-G6AB-CUP1 plasmid.Wherein, swimming lane 1 is with AY-510B24M
The GAL6A segment gone out for template amplification；Swimming lane 2 is the GAL6A segment that amplifies using pUC-G6AB-CUP1 plasmid as template；Swimming
Road 3 is the GAL6B segment gone out by template amplification of AY-510B24M；Swimming lane 4 is to expand using pUC-G6AB-CUP1 plasmid as template
Increase GAL6B segment out；Swimming lane 5 is the CUP1 segment gone out by template amplification of Yep-C；Swimming lane 6 is with pUC-G6AB-CUP1 plasmid
For template, the CUP1 segment amplified；Swimming lane 7 is to amplify the GAL6A- come using pUC-G6AB-CUP1 plasmid as template
CUP1-GAL6B genetic fragment, swimming lane M are DL5000DNA marker.
4) GAL6A-CUP1-GAL6B homologous recombination segment is imported by Lac using lithium acetate transformation method+The a of saccharomyces cerevisiae
Type and α type monoploid, the genes of brewing yeast engineering haploid strains after obtaining homologous recombination；Lac used in the present embodiment+Wine brewing
Yeast be specially saccharomyces cerevisiae AY-510B24M disclosed in patent CN102604849B (Saccharomyces cerevisiae,
Deposit number is CGMCC No 5843)
Fig. 3 is a and α type positive restructuring monoploid PCR verification result.Wherein swimming lane 1 is to amplify in a type recombination monoploid
The GAL6A-CUP1-GAL6B genetic fragment come；Swimming lane 2 is that α type recombinates the GAL6A-CUP1-GAL6B for amplifying and in monoploid
Genetic fragment；Swimming lane 3 is that AY-510B24M is template, the complete GAL6 segment come out using identical primer amplification；Swimming lane 4 is
DL5000 DNA marker.Swimming lane 1,2 shows the GAL6 gene in parent by GAL6A-CUP1-GAL6B homologous recombination piece
Section substitution.
5) by after purification saccharomyces cerevisiae a type and α type recombination monoploid merge, pass through resistant panel and the experiment of raw spore
Screening obtains can use the engineering bacteria (amphiploid) of whey.
(2) specific sequence of engineering strain
Contain one section of specific sequence in the engineering strain AY5MG chromosome of acquisition, after being sequenced by PCR amplification
Carry out the identification of negative regulator gene GAL6 removal.
The primer sequence of specific fragment amplification is respectively as follows:
G6A-U:5'-AAAGAATTCGCGGAAAGGCAGGCAATA-3’
G6B-D:5'-ATCAAGCTTATGGTAGCCGAATGAATGAAAT-3’
The gene order of specific fragment is shown in sequence table 1.
Embodiment 2: the research of fuel ethanol project bacterium fermenting property is produced using whey
AY5MG and its parent AY-510B24M are respectively connected in 20mL glucose culture solution, 30 DEG C are incubated overnight 12h；
Bacterium solution is all gone in 200mL whey medium after centrifuge washing, 30 DEG C of stationary culture fermentations.Whey medium is (g/L):
Whey powder 120 (lactose content 53.1g/L), (NH4)2SO45, MgSO4·7H2O 1 is settled to 1L with water.It is every during fermentation
Every concussion sampling for 24 hours, and record weightlessness；After fermentation, stop cultivating and weighing；Measure remaining sugar concentration, the alcohol of fermentation liquid
Concentration and dry cell weight characterize its comprehensive performance, the results are shown in Table 2.
2 saccharomyces cerevisiae recipient bacterium of table and engineering bacteria fermenting property in whey
Note: shown data are the average value of three parallel test results.
Embodiment 3: whey is existing using the glucose repression of type saccharomyces cerevisiae engineered yeast and starting strain whey decomposition product
As research
Engineering bacteria and recipient bacterium are respectively connected in 5mL YEPD culture solution, 30 DEG C are incubated overnight 12h；Bacterium solution is all turned
Into 20mL galactolipin culture solution, 30 DEG C of cultures are for 24 hours.Prepare simulation whey decomposition product culture medium: glucose 3g, galactolipin
3g, (NH4)2SO40.5g, MgSO4·7H2O 0.1g, yeast powder 0.2g, peptone 0.1g, KH2PO40.3g, distilled water
100mL.It is inoculated with by 10% inoculum concentration, 30 DEG C of stationary cultures.Oscillation sampling at regular intervals, it is dense to survey different sugar during fermentation
Degree, is as a result shown in Fig. 5.Lactose in Whey first has to resolve into glucose and galactolipin in saccharomyces cerevisiae engineered yeast strain, then again
Into respective metabolic pathway.However for saccharomyces cerevisiae, the presence of glucose can strictly inhibit the utilization of galactolipin, this
It is exactly glucose repression phenomenon.From fig. 5, it can be seen that removal GAL6 gene, can also slow down the glucose repression of AY51024M
Phenomenon shortens the fermentation period in muscovado.
Embodiment 4: gained engineering bacteria is compared with recipient bacterium patience
Engineering bacteria and recipient bacterium are connected to progress level-one culture, 30 DEG C of stationary cultures in 6mL/ pipe YEPD respectively to take afterwards for 24 hours
Seed liquor 0.5mL is connected in the YEPD culture medium of different alcohol concentrations, and 30 DEG C of stationary culture observations produce gas situation.Temperature tolerance is real
It tests to take seed liquor 0.5mL to be connected in YEPD culture medium, is cultivated in different temperatures, observe it and produce gas situation.Alcohol tolerance and
Temperature tolerance is shown in Table 4, and specific medium component content is shown in Table 3.
The different alcohol concentration medium component contents of table 3
4 engineering bacteria of table is compared with recipient bacterium alcohol tolerance, temperature tolerance
Note: "+" is growth, and "-" is not grow；Shown result is three parallel laboratory test average results
Sequence table
<110>Hebei Science & Technology Normal College
<120>a kind of method using whey production ethyl alcohol
<130> 1
<141> 2018-10-19
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2948
<212> DNA
<213>saccharomyces cerevisiae (Saccharomyces cerevisiae)
<220>
<221> variation
<222> (1…2948)
<400> 1
aggagcaagc aactgaccca atattgactg ccactggacc tgaagacatg caacaaagtg 60
gaattcgcgg aaaggcaggc aatatgcttt acaaacaggc aaattttgtc tcttaatcct 120
gtgaagaggc atcgtatatt cagtgctttt tgaaatttag aaaatagtcg ctttattgat 180
tatgctacat atgttatttt gctttagtat gtgtcaaata gcatgcaatc gccgggtagt 240
gagtgacaca tccttttttc ccctcccacc agaaagtcac ctgctctctt aatcaatggg 300
gttgcatgtg gtaatcaatt gataacgccg gctgacaagt cgccgacggg cacccataaa 360
taaacgataa atagcccttg ctcttttgtt acatgcttcc tacttctgtt tcccggtcct 420
tgtatctcaa gacgtttaga agccaccttt tacgagcacc acaaattgtt ttaaaaagaa 480
tgtcctcttc catcgatatc agtaagatca actcttggaa caaagagttt caatccgact 540
taacccatca attggcaacc actgtcctta agaattataa tgccgatgat gcactgttga 600
acaagactag actgcaaaag caagataaca gggttttcaa cactgttgtc tctactgatt 660
ccactccagt caccaaccaa aaaagctctg gtagatgttg gttgtttgcc gcggtaccca 720
gctgaagctt cgtacgctgc aggtcgacaa cccttaatat aacttcgtat aatgtatgct 780
atacgaagtt attaggtcta gagatctgtt tagcttgcct cgtccccgcc gggtcacccg 840
gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg 900
gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca 960
tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc 1020
tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg 1080
tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt 1140
aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg 1200
aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt 1260
ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga 1320
agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta 1380
cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc 1440
attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag 1500
cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag 1560
tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg 1620
tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt 1680
ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt 1740
tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt 1800
ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat 1860
accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac 1920
ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga 1980
tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt 2040
gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat 2100
ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt 2160
aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta 2220
acgccgccat ccagtgtcga aaacgagctc tcgagaaccc ttaatataac ttcgtataat 2280
gtatgctata cgaagttatt aggtgatatc agatccacta gtggcctatg cggatccctg 2340
ccacgtcgat gaaacgtcta aattaccact tcgctaccgc gttgaaaatt cctggggtaa 2400
agactccggt aaagacggat tatacgtgat gactcaaaag tacttcgagg agtactgctt 2460
tcaaattgtg gtcgatatca atgaattgcc aaaagagctg gcttcaaaat tcacctcagg 2520
taaggaagag ccgattgtct tgcccatctg ggacccaatg ggtgctttgg ccaaataaat 2580
agtttcagca gctctgatgt agatacacgt atctcgacat gttttatttt tactatacat 2640
acataaaaga aataaaaaat gataacgtgt atattattat tcatataatc aatgagggtc 2700
acaaaggaaa ggttagcata ttaaataact gagctgatac ttcaacagca tcgctgaaga 2760
gaacagtatt gaaaccgaaa cattttctaa aggcaaacaa ggtactccat atttgctgga 2820
cgtgttcttt ctctcgtttc atatgcataa ttctgtcata agcctgttct ttttcctggc 2880
ttaaacatcc cgttttgtaa aagagaaatc tattccacat atttcattca ttcggctacc 2940
ataagctt 2948
Claims (3)
1. a kind of method using whey production ethyl alcohol, which is characterized in that it is CGMCC that the method, which is using deposit number,
The Wine brewing yeast strain of No.11223 stationary culture fermentation next life producing and ethanol in the culture medium containing whey or lactose, fermentation week
Phase 54h.
2. the method as described in claim 1, which is characterized in that the composition of the culture medium are as follows: whey powder 120g/L or lactose
53.1g/L, (NH4)2SO45g/L, MgSO4·7H2O 1g/L, solvent are water.
3. method according to claim 1 or 2, which is characterized in that the temperature of the fermentation is 30 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811220905.4A CN109161565B (en) | 2015-09-29 | 2015-09-29 | Method for producing ethanol by using whey |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510632088.3A CN105199975B (en) | 2015-09-29 | 2015-09-29 | One plant height effect utilizes the saccharomyces cerevisiae engineered yeast and its construction method of whey producing and ethanol |
| CN201811220905.4A CN109161565B (en) | 2015-09-29 | 2015-09-29 | Method for producing ethanol by using whey |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510632088.3A Division CN105199975B (en) | 2015-09-29 | 2015-09-29 | One plant height effect utilizes the saccharomyces cerevisiae engineered yeast and its construction method of whey producing and ethanol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109161565A true CN109161565A (en) | 2019-01-08 |
| CN109161565B CN109161565B (en) | 2021-06-18 |
Family
ID=54947938
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510632088.3A Active CN105199975B (en) | 2015-09-29 | 2015-09-29 | One plant height effect utilizes the saccharomyces cerevisiae engineered yeast and its construction method of whey producing and ethanol |
| CN201811220905.4A Active CN109161565B (en) | 2015-09-29 | 2015-09-29 | Method for producing ethanol by using whey |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510632088.3A Active CN105199975B (en) | 2015-09-29 | 2015-09-29 | One plant height effect utilizes the saccharomyces cerevisiae engineered yeast and its construction method of whey producing and ethanol |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN105199975B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115806889B (en) * | 2022-09-28 | 2024-06-11 | 山东大学 | Saccharomyces cerevisiae engineering bacteria for improving gene expression level and construction method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20100048224A (en) * | 2008-10-30 | 2010-05-11 | 삼성전자주식회사 | Gene increasing metabolic availability of galactose, recombinant vector and recombinant microorganism containing the same |
| CN102604849A (en) * | 2012-03-23 | 2012-07-25 | 天津科技大学 | Saccharomyces cerevisiae engineering bacterial strain capable of efficiently using lactose to produce fuel ethanol |
-
2015
- 2015-09-29 CN CN201510632088.3A patent/CN105199975B/en active Active
- 2015-09-29 CN CN201811220905.4A patent/CN109161565B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20100048224A (en) * | 2008-10-30 | 2010-05-11 | 삼성전자주식회사 | Gene increasing metabolic availability of galactose, recombinant vector and recombinant microorganism containing the same |
| CN102604849A (en) * | 2012-03-23 | 2012-07-25 | 天津科技大学 | Saccharomyces cerevisiae engineering bacterial strain capable of efficiently using lactose to produce fuel ethanol |
Non-Patent Citations (3)
| Title |
|---|
| S OSTERGAARD 等: "Increasing galactose consumption by saccharomyces cerevisiase through metabolic engineering of the GAL gene regulatory network", 《NATURE BIOTECHNOL》 * |
| 邹静 等: "Lac+酿酒酵母工程菌中GAL80基因敲除对乳糖利用的影响", 《酿酒科技》 * |
| 阎隆飞 等: "《分子生物学》", 30 September 1993, 北京农业大学出版社 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105199975B (en) | 2018-12-07 |
| CN109161565B (en) | 2021-06-18 |
| CN105199975A (en) | 2015-12-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mo et al. | Kluyveromyces marxianus developing ethanol tolerance during adaptive evolution with significant improvements of multiple pathways | |
| Wisselink et al. | Novel evolutionary engineering approach for accelerated utilization of glucose, xylose, and arabinose mixtures by engineered Saccharomyces cerevisiae strains | |
| Lee et al. | Kinetic studies on a highly productive strain of Zymomonas mobilis | |
| Du Preez et al. | A quantitative screening of some xylose-fermenting yeast isolates | |
| CN103201376A (en) | Yeast production culture for the production of butanol | |
| JPS6357039B2 (en) | ||
| JP6189941B2 (en) | Yeast strain capable of metabolizing xylose and resistant to inhibitors, method for obtaining the yeast strain and use thereof | |
| CN101983240B (en) | Flocculent yeast and method for production thereof | |
| CN103849576A (en) | Recombined saccharomyces cerevisiae strain with stress tolerance | |
| CN104561076B (en) | A kind of high yield L serines recombinate the structure and its fermentation process of Corynebacterium glutamicum | |
| CN100558882C (en) | A kind of wood sugar and glucose production alcoholic acid recombinant Saccharomyces cerevisiae of utilizing | |
| CN102604849B (en) | Saccharomyces cerevisiae engineering bacterial strain capable of efficiently using lactose to produce fuel ethanol | |
| CN103373901B (en) | Method for extracting erythritol from erythritol mother liquor and special barm strain for erythritol | |
| CN106414735B (en) | Optimized propagation pentose fermentation strain | |
| CN105199975B (en) | One plant height effect utilizes the saccharomyces cerevisiae engineered yeast and its construction method of whey producing and ethanol | |
| CN111334442A (en) | High-temperature-resistant saccharomyces cerevisiae strain and application thereof | |
| CN103820347A (en) | Industrial saccharomyces cerevisiae strain with acetic acid tolerance | |
| CN104099377B (en) | A kind of method for improving Alcohol Production efficiency | |
| Corbin et al. | Low-input fermentations of Agave tequilana leaf juice generate high returns on ethanol yields | |
| CN107974453A (en) | It is a kind of to improve brewing yeast cell vigor and the methods and applications of multiple resistance | |
| CN104403956B (en) | The structure of xylitol high temperature high-yielding engineering bacterial strain and application | |
| Tyurin et al. | Synthetic 2, 3-Butanediol Pathway Integrated Using Tn 7-tool and Powered Via Elimination of Sporulation and Acetate Production in Acetogen Biocatalyst | |
| CN105255952A (en) | Method for improving ethyl alcohol production efficiency through INO2 gene overexpression | |
| Amin et al. | Comparative study of D-xylose conversion to ethanol by immobilized growing or non-growing cells of the yeast Pachysolen tannophilus | |
| CN116716321B (en) | Application of HMX1 and encoding gene thereof in improving fermentation performance and acetic acid tolerance of saccharomyces cerevisiae xylose |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |