CN105368729B - One plant of acidproof flocculability an industrial strain of S.cerevisiae strain and construction method - Google Patents
One plant of acidproof flocculability an industrial strain of S.cerevisiae strain and construction method Download PDFInfo
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Abstract
Disclosed by the invention is one plant of acidproof flocculability an industrial strain of S.cerevisiae strain and construction method, and amphilepsis character can only effectively be had concurrently mostly by solving the bacterial strain that existing integration technology merges out, and cannot effectively improve fusion after bacterial strain inhereditary feature performance the problem of.The present invention include the saccharomyces cerevisiae (Saccharomyces cerevisiae SEB4) bacterial strain is preserved in China General Microbiological culture presevation administrative center, deposit number CGMCC11324.The present invention also provides the construction methods of Saccharomyces Cerevisiae in S EB4.Saccharomyces Cerevisiae in S EB4 bacterial strain after merging in the present invention, not only has amphilepsis character concurrently, has both had the sub No. 1 good flocculability of parent fusion and fermentability and the good acid-fast ability of parent SEB2；And it is superior to parent strain on ethanol production and glucose consumption rate.
Description
Technical field
The present invention relates to a kind of yeast strains, and in particular to be one plant of acidproof flocculability an industrial strain of S.cerevisiae strain and
The construction method of the bacterial strain.
Background technique
Bio-fuel ethyl alcohol is considered as the important energy source of alternative conventional fuel oil.Saccharomyces cerevisiae is that bio-ethanol industry is raw
Main microorganism to be used in production, the optimum pH of general bacterial strain are 4-5.When pH is higher or lower than the range, the life of bacterial strain
The decline of long and fermentability.And large-scale industrial production needs control living contaminants well, reduces fermentation system as far as possible
PH value is relatively effective measure.In addition, lignocellulose-like biomass is important fuel ethanol production raw material, it is utilized
First step need to be pre-processed using diluted acid, resulting hydrolyzate pH value is lower, needs to be adjusted to suitable pH value
It could be used for the alcohol fermentation of yeast afterwards.Therefore, the quality yeast bacterial strain for obtaining tolerance low ph value helps to improve ethyl alcohol production
Stability, reduce production cost.
Protoplast fusion is a kind of common Microbial Breeding technology, refers to through artificial method, makes inhereditary feature not
The protoplast of two same cells is merged, and has the process of the stabilization recon of amphilepsis character concurrently so as to obtaining.With
Other breeding techniques are compared, Protoplast Fusion Technique have recombination frequency it is higher, by maqting type or cause educate type limited it is smaller and
More complete etc. advantages of inhereditary material, thus be widely applied by domestic and international Microbial Breeding scholar.
Summary of the invention
It is an object of the invention to solve the bacterial strain that existing integration technology merges out can only effectively have amphilepsis concurrently mostly
Character, and cannot effectively improve fusion after bacterial strain inhereditary feature performance the problem of；Amphilepsis can effectively be had concurrently by providing one plant
Character, the acidproof flocculability work that parent strain can also be superior on ethanol production, glucose utilization and wear rate simultaneously
Industry Wine brewing yeast strain, and provide the construction method of the bacterial strain.
In order to achieve the above objectives, the specific technical solution of the present invention is as follows:
One plant of acidproof flocculability an industrial strain of S.cerevisiae strain, the saccharomyces cerevisiae (Saccharomyces cerevisiaeSEB4) bacterial strain is preserved in China General Microbiological culture presevation administrative center, deposit number CGMCC11324,
Time is on September 6th, 2105.
Fused Saccharomyces Cerevisiae in S EB4 bacterial strain, not only has amphilepsis character concurrently in the present invention, that is, has parent fusion
The good flocculability of sub- RHZ-1 and fermentability and the good acid-fast ability of parent SEB2；And it is in ethanol production, grape
Parent strain is superior on sugar consumption amount and wear rate.
It was proved that 30 °C, pH 2.2, enter sugared concentration be 150 g/l under the conditions of:
When 24 h, Saccharomyces Cerevisiae in S EB4 bacterial strain sugar consumption amount is 78.7%, sugar consumption rate is 4.9 g/l/h, ethanol production
For 47.87 g/l, the sub- RHZ-1 sugar consumption amount of parent fusion is 64.9%, sugar consumption rate is 4.1 g/l/h, ethanol production is
35.93 g/l, parent's SEB2 sugar consumption amount is 71.6%, sugar consumption rate is 4.5 g/l/h, ethanol production is 32.69 g/l；
When 48 h, all bacterial strains can all consume glucose, and Saccharomyces Cerevisiae in S EB4 bacterial strain ethanol production is 57.76
G/l, the sub- RHZ-1 ethanol production of parent fusion are 51.93 g/l, and parent's SEB2 ethanol production is 54.42 g/l.
The construction method of Wine brewing yeast strain as described in claim 1, comprising the following steps:
1) building of auxotrophic mutation bacterium
1.1) with saccharomyces cerevisiaeSaccharomyces cerevisiae IR-2 and saccharomyces cerevisiaeSaccharomyces cerevisiae SEB1 is parent, obtains fusant RHZ-1 by cell fusion mode；
Wherein, SEB1 bacterial strain has ethanol production, is preserved in China General Microbiological culture presevation administrative center,
Deposit number is CGMCC11321.IR-2 bacterial strain: being isolated from Indonesia's fermented food, has ethanol production, has
Flocculability, source are recorded in the following references: Hiroshi K, Yoshio S, Toshio M, Harumi K, Yorikazu
S. 1985. Continuous ethanol fermentation with cell recycling using
flocculating yeast. J. Ferment. Technol. 63:159-165；Title after document translation are as follows: use
Flocculability yeast carries out the continuous alcohol fermentation with cell cycle；Above-mentioned IR-2 biomaterial is from Japanese industries Technical Integration Studies
Institute (National Institute of Advanced Industrial Science and Technology, AIST) obtains
?.The fusion method of above-mentioned IR-2 bacterial strain and SEB1 bacterial strain is as follows:
SEB1 and IR-2 is grown 3 days on producing Spore cultivation base (0.5% lithium acetate, 2% agar), and resulting ascus is through yeast
Cracking enzymatic treatment obtains spore, handles 2 hours through ethylmethane sulfonate (EMS), YPD plate culture is coated onto after ultrasonic disperse, so
It is grown 4 days in photocopy to minimum nutrition plate afterwards, examines the auxotroph of the non-growing bacterium colony on minimum nutrition plate.From
SEB1 obtains the bacterial strain SIV-2 of isoleucine and valine deficiency, obtains the bacterial strain IL-1 of lysine deficient from IR-2.
The composition of above-mentioned YPD plate are as follows: 2% glucose, 1% yeast powder, 2% polyprotein peptone, 2% agar, remaining is distillation
Water；The composition of above-mentioned minimum nutrition plate are as follows: 2% glucose, 0.67% without amino acid yeast nitrogen, and 2% agar, remaining is distillation
Water.
After yeast cracks enzymatic treatment 2 hours, obtained protoplast mixes SIV-2 and IL-1 in 30% PEG6000
Processing 15 minutes is coated on regenerated plate and selection plate.The bacterium colony grown on selection plate is considered as fusant, merges subnumber
Amount is fusion efficiencies with plasmic quantity ratio is regenerated.Obtaining 8 plants of fusant RHZ-1(fusion efficiencies altogether is 1.1 × 10-5),
Screening obtains the fusant using 25% molasses fermented producing and ethanol most fast (1.3 g/l/h) and concentration of alcohol highest (65 g/l)
RHZ-1。
The composition of above-mentioned regenerated plate are as follows: 2% glucose, 0.5% yeast powder, 1% polyprotein peptone, 4.5% potassium chloride, 2% fine jade
Rouge, remaining is distilled water；The composition of above-mentioned selection plate are as follows: 2% glucose, 0.67% without amino acid yeast nitrogen, 2% agar,
Remaining is distilled water.
1.2) fusant RHZ-1 and SEB2 are subjected to ultraviolet mutagenesis, filter out the single of fusant RHZ-1 and SEB2 respectively
Auxotrophic mutation bacterium；SEB2 is deposited in China General Microbiological culture presevation administrative center, and deposit number is
CGMCC11322。
1.3) the good bacterial strain of flocculability is filtered out from fusant RHZ-1 single auxotrophic mutation bacterium, and to this
Bacterial strain carries out growth ability and fermenting property evaluation experimental, picks out the optimal bacterial strain of fermenting property and melts as subsequent native plastid
The auxotrophic mutation bacterium KFU of conjunction；Growth ability and fermenting property evaluation are carried out to SEB2 single auxotrophic mutation bacterium
Experiment, filters out auxotrophic mutation bacterium KAGOU of the bacterial strain as subsequent native plast fusion of best performance；
2) building of Protoplast fusant
2.1) using auxotrophic mutation bacterium KFU and auxotrophic mutation bacterium KAGOU as parent, protoplast is carried out
The fusion of cell obtains fusant RHZ-2；
2.2) by fused suspension to plate, the fusant RHZ-2 grown is screened, is filtered out same
When there is flocculability and acid-fast ability and the optimal bacterial strain of fermenting property, the bacterial strain are Saccharomyces Cerevisiae in S EB4.
The present invention, in conjunction with ultraviolet mutagenesis and Protoplast Fusion Technique, is obtained using 2 industrial saccharomyces cerevisiaes as parent strain
The preferably acidproof flocculability yeast strain of 1 plant of fermenting property, can provide excellent microbial resources for the production of alcohol fuel.
The bacterial strain that method of the invention constructs not only has amphilepsis character concurrently, that is, has the good flocculation of the sub- RHZ-1 of parent fusion
Property and fermentability and the good acid-fast ability of parent SEB2；And bacterial strain inhereditary feature performance after fusion can also be effectively improved,
The fusant bacterial strain of method building i.e. of the invention is superior to parent on ethanol production, glucose utilization and glucose consumption rate
This bacterial strain.
Further, the specific operation process of the ultraviolet mutagenesis is as follows:
Fusant RHZ-1 and SEB2 are transferred to respectively in 2 % YPD culture mediums, 30 °C of 16 h of activation；Bacterium solution is through sterile water
It is applied to after dilution on YPD plate, after plate irradiates 50 s in the UV lamp, in 30 °C of stationary cultures.
Further, the screening process of the single auxotrophic mutation bacterium is as follows:
The bacterial strain of the YPD plate culture handled through ultraviolet lamp is fallen in photocopy to MM plate, and is incubated overnight in 30 °C, it is right
Than the bacterium colony that YPD and MM plate is grown, picking out from YPD plate can not be in the bacterium colony of MM plated growth, i.e. auxotroph
Mutant bacteria；
Different nutriments is added to MM plate, a kind of nutrients of each plate connects auxotrophic mutation bacterium simultaneously
It is cultivated in kind to YPD and MM plate, identifies its auxotrophy type, and filter out single auxotrophic mutation bacterium.
Wherein, 2%YPD culture medium includes: 20 g/l glucose, 10 g/l yeast extract powders, 20 g/l peptones, remaining
For distilled water；MM culture medium includes: 1.7 g/l without amino yeast nitrogen, 10 g/l yeast powders, 20 g/l glucose, remaining is
Distilled water.Plate is added to the agar of 20 g/l on the basis of culture medium.
Compared with prior art, the present invention have the following advantages that and the utility model has the advantages that
Saccharomyces Cerevisiae in S EB4 bacterial strain after merging in the present invention, not only has amphilepsis character concurrently, that is, has parent fusion
The good flocculability of sub- RHZ-1 and fermentability and the good acid-fast ability of parent SEB2；And it is in ethanol production, grape
Parent strain is superior on sugar consumption amount and glucose consumption rate.
The preservation information of the microorganism of preservation involved in the present invention is as follows:
Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center；Address: Chaoyang District, Beijing City
No. 3 Institute of Microorganism, Academia Sinica of institute of North Star West Road 1；Preservation date: on September 6th, 2015；Deposit number:
CGMCC11321,CGMCC11322,CGMCC11324；Classification naming: Saccharomyces cerevisiae.
Detailed description of the invention
The growth result schematic diagram of auxotrophic mutation bacterium when Fig. 1 is addition different nutriment.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, embodiments of the present invention are not limited thereto.
Embodiment 1
One plant of acidproof flocculability an industrial strain of S.cerevisiae strain, the saccharomyces cerevisiae (Saccharomyces cerevisiae
SEB4) bacterial strain is preserved in China General Microbiological culture presevation administrative center, deposit number CGMCC11324.The saccharomyces cerevisiae
The specific construction method of bacterial strain SEB4 is as follows:
1) building of auxotrophic mutation bacterium
1.1) with saccharomyces cerevisiaeSaccharomyces cerevisiaeIR-2 and saccharomyces cerevisiaeSaccharomyces cerevisiae SEB1 is parent, obtains fusant RHZ-1 by cell fusion mode.
The SEB1 bacterial strain in the present embodiment has ethanol production, is preserved in China General Microbiological culture presevation pipe
Reason center, deposit number CGMCC11321.IR-2 bacterial strain: being isolated from an Indonesian fermented food, has ethyl alcohol
Production capacity has flocculability, and source is recorded in the following references: Hiroshi K, Yoshio S, Toshio M, Harumi
K, Yorikazu S. 1985. Continuous ethanol fermentation with cell recycling
using flocculating yeast. J. Ferment. Technol. 63:159–165；Title after document translation
Are as follows: the continuous alcohol fermentation with cell cycle is carried out using flocculability yeast；Above-mentioned IR-2 biomaterial is from Japanese industries technology
Comprehensive study institute (National Institute of Advanced Industrial Science and Technology,
AIST it) obtains.The fusion method of above-mentioned IR-2 bacterial strain and SEB1 bacterial strain is as follows:
SEB1 and IR-2 is grown 3 days on producing Spore cultivation base, and resulting ascus obtains spore through yeast cracking enzymatic treatment,
It is handled 2 hours through ethylmethane sulfonate (EMS), YPD plate culture is coated onto after ultrasonic disperse, then photocopy to minimum nutrition plate
Upper growth 4 days, examines the auxotroph of the non-growing bacterium colony on minimum nutrition plate.Isoleucine and figured silk fabrics are obtained from SEB1
The bacterial strain SIV-2 of propylhomoserin deficiency obtains the bacterial strain IL-1 of lysine deficient from IR-2.
After yeast cracks enzymatic treatment 2 hours, obtained protoplast mixes SIV-2 and IL-1 in 30% PEG6000
Processing 15 minutes is coated on regenerated plate and selection plate.The bacterium colony grown on selection plate is considered as fusant, merges subnumber
Amount is fusion efficiencies with plasmic quantity ratio is regenerated.Obtaining 8 plants of fusant RHZ-1(fusion efficiencies altogether is 1.1 × 10-5),
Screening obtains the fusant using 25% molasses fermented producing and ethanol most fast (1.3 g/l/h) and concentration of alcohol highest (65 g/l)
RHZ-1。
The composition of above-mentioned production Spore cultivation base are as follows: 0.5% lithium acetate, 2% agar, remaining is distilled water；Above-mentioned YPD plate
It constitutes are as follows: 2% glucose, 1% yeast powder, 2% polyprotein peptone, 2% agar, remaining is distilled water；Above-mentioned minimum nutrition plate
It constitutes are as follows: 2% glucose, 0.67% without amino acid yeast nitrogen, and 2% agar, remaining is distilled water.
The composition of above-mentioned regenerated plate are as follows: 2% glucose, 0.5% yeast powder, 1% polyprotein peptone, 4.5% potassium chloride, 2% fine jade
Rouge, remaining is distilled water；The composition of above-mentioned selection plate are as follows: 2% glucose, 0.67% without amino acid yeast nitrogen, 2% agar,
Remaining is distilled water.
1.2) fusant RHZ-1 and SEB2 are subjected to ultraviolet mutagenesis, filter out the single of fusant RHZ-1 and SEB2 respectively
Auxotrophic mutation bacterium；SEB2 is deposited in China General Microbiological culture presevation administrative center, and deposit number is
CGMCC11322.Detailed process is as follows:
Firstly, fusant RHZ-1 and SEB2 are transferred to respectively in 2% YPD culture medium, 30 °C of 16 h of activation；Bacterium solution is through nothing
It is applied on YPD plate after the dilution of bacterium water, after plate irradiates 50 s in the UV lamp, in 30 °C of stationary cultures.
Secondly, the bacterial colony photographic reprinting for growing plate after 2 d of stationary culture is to MM plate, and it is incubated overnight in 30 °C.It is right
Than the bacterium colony that YPD and MM plate is grown, picking out from YPD plate can not be in the bacterium colony of MM plated growth, i.e. auxotroph
Mutant bacteria.It is verified by 4 repeated screenings, finally obtains the auxotrophic strain of 3 plants of fusant RHZ-1, every plant is selected 2
Clone, is respectively designated as: KFU-A1, KFU-A2；KFU-B1,KFU-B2；KFU-C1,KFU-C2；The nutrition for obtaining 5 plants of SEB2 lacks
Swaged bacterial strain, every plant is selected 2 clones, is respectively designated as: KAGOU-A1, KAGOU-A2；KAGOU-B1,KAGOU -B2；
KAGOU-C1,KAGOU -C2；KAGOU-D1,KAGOU-D2；KAGOU-E1,KAGOU-E2.
Finally, different nutriments is added into MM plate, and a kind of nutrients of each plate, totally 22 kinds, as shown in table 1,
Auxotrophic mutation bacterium is inoculated on YPD and MM plate simultaneously and is cultivated, cultivation results are as shown in Figure 1, be to melt in Fig. 1 (a)
The auxotrophic strain of zygote RHZ-1 is the auxotrophic strain of SEB2 in Fig. 1 (b).
Shown by Fig. 1: KFU-A1, KFU-A2 and KAGOU-D1, KAGOU-D2 are 4 plants of uracil-deficient bacterial strains
(Ura-)；KFU-B1, KFU-B2 are 2 plants of lysine deficient bacterial strain (Lys-)；KFU-C1, KFU-C2 are 2 plants of cysteine defects
Type bacterial strain (Cys-)；KAGOU-B1, KAGOU-B2 are 2 plants of Adenine nucleotides bacterial strain (Ade-)；KAGOU-E1, KAGOU-E2 2
Strain histidine deficient bacterial strain (His-)；KAGOU-A1, KAGOU-A2, KAGOU-C1, KAGOU-C2 are equal on all MM plates
It does not grow.
Shown by the above results: this 4 plants of bacterium of KAGOU-A1, KAGOU-A2, KAGOU-C1 and KAGOU-C2 be two kinds or
A variety of Different Nutrition deficient strains are not used to the screening of Protoplast Fusant, subsequent no longer to investigate to it.
The single auxotrophic mutation bacterium that the present embodiment filters out are as follows: KFU-A1, KFU-A2, KFU-B1, KFU-B2,
KFU-C1 and KFU-C2；And KAGOU-B1, KAGOU-B2, KAGOU-D1, KAGOU-D2, KAGOU-E1 and KAGOU-E2.
Table 1
Wherein, 2%YPD culture medium: 20 g/l glucose, 10 g/l yeast extract powders, 20 g/l peptones.MM culture medium:
1.7 g/l are without amino yeast nitrogen, 10 g/l yeast powders, 20 g/l glucose.Plate is added on the basis of culture medium
The agar of 20 g/l.
1.3) parent strain as subsequent native plast fusion is filtered out
1.3.1 the good bacterial strain of flocculability in fusant RHZ-1 single auxotrophic mutation bacterium) is filtered out, and to this
Bacterial strain carries out growth ability and fermenting property evaluation experimental, picks out the optimal bacterial strain of fermenting property and melts as subsequent native plastid
The auxotrophic mutation bacterium KFU of conjunction；Detailed process is as follows:
Fusant RHZ-1 single auxotrophic mutation bacterium is seeded in 2%YPDU culture medium and cultivates 16 h, takes 5 respectively
Ml bacterial culture fluid is uniform through vortex oscillation in test tube, stands the flocculability of observation bacterial strain after different time, sends out by observation
Now in addition to bacterial strain KFU-A2, other bacterial strains remain with the good flocculability of the sub- RHZ-1 of parent fusion.The 2%YPDU culture medium
It include: 2 g/l peptones, 1 g/l ammonium sulfate, 20 g/l glucose, 0.04 g/l uracil, remaining is distilled water.
Therefore, the growth ability of all bacterial strains in acid condition in addition to bacterial strain KFU-A2 is investigated；Growth ability evaluation step
Suddenly are as follows: be seeded to fusant RHZ-1 mutant strain respectively in the 2%YPDU culture medium of pH 3.0 and pH 2.7, and in 35 °C of items
The culture in small test tubes culture instrument (ADVANTEC, Japan) under part, it is small that instrument automatically records different pH value hypothallus growths 24
When OD660Value；The results are shown in Table 2.
Table 2
By table 2 it is found that bacterial strain KFU-A1, KFU-B1, KFU-B2 are under the conditions of pH 3.0 and 2.7 pH, growth ability is equal
It is close with the sub- RHZ-1 bacterial strain of parent fusion；The growth of bacterial strain KFU-C1 and KFU-C2 has a part inhibition, but KFU-C1 is grown in
It is seriously suppressed under the conditions of pH 2.7.
Therefore, selection bacterial strain KFU-A1, KFU-B2 and KFU-C2 carries out fermenting property evaluation.
The specific evaluation procedure of fermenting property evaluation is as follows: by the single auxotrophic mutation of the sub- RHZ-1 of parent fusion
Bacterial strain is seeded to respectively in 5%YPDU culture medium (pH natural), and in 30 °C, 160 rpm (shaking table oscillation) preculture, 16 h.Again
It is seeded in the 15%YPDU culture medium of pH 2.7, in 35 °C, 400 rpm (magnetic agitation), 48 h of fermentation periodically take fermentation liquid to examine
Survey total cell number, concentration of glucose and concentration of alcohol.Following result: bacterial strain KFU-B2 is obtained after evaluating by fermenting property
(Lys-) fermenting property with higher, in cell survivaling number, ethanol production, glucose consumption aspect is better than other bacterial strains.
The 5%YPDU culture medium: the concentration of glucose in 2%YPDU culture medium is adjusted to 50 g/l, remaining component is constant；It is described
15% YPDU culture medium: the concentration of glucose in 2%YPDU culture medium is adjusted to 150 g/l, remaining component is constant.
To sum up, thus select KFU-B2 as the parent strain of subsequent native plast fusion.
1.3.2 growth ability and fermenting property evaluation experimental) are carried out to SEB2 single auxotrophic mutation bacterium, filtered out
Auxotrophic mutation bacterium KAGOU of the bacterial strain of best performance as subsequent native plast fusion；Detailed process is as follows:
Using parent strain SEB2 as control, is evaluated and walked using growth ability identical with above-mentioned fusant RHZ-1 bacterial strain
Suddenly, compare the growth rate μ of each mutant strain of SEB2max(h-1), OD660Reach 0.5 required time tOD660=0.5(min) it and reaches
The maximum OD arrivedmax, concrete outcome is as shown in table 3.
Table 3
By table 3: under the conditions of pH 3.0 and 2.7 pH, the growth ability of bacterial strain KAGOU-E1, KAGOU-E2 with
Parent SEB2 is similar.And the growth of other bacterial strains is centainly inhibited, and when pH is 2.7, inhibits situation more obvious.
Meanwhile bacterial strain KAGOU-B2, KAGOU-D2 and KAGOU-E2 being selected to carry out fermenting property evaluation.I.e. by with it is upper
The identical fermenting property evaluation procedure of fusant RHZ-1 bacterial strain is stated, is compared with parent strain SEB2, fermenting experiment is shown:
KAGOU-E2 (His-) at cell survivaling number, ethanol production, glucose consumption aspect, than other two bacterial strain with preferable
Performance.
To sum up, therefore select KAGOU-E2 as the parent strain of subsequent native plast fusion.
2) building of Protoplast fusant
2.1) with auxotrophic mutation bacterium KFU-B2 (Lys-) and auxotrophic mutation bacterium KAGOU-E2 (His-) conduct
Parent carries out the fusion of protoplasm somatocyte, obtains fusant RHZ-2；
In protoplast preparation process, measure two bacterial strains protoplast formation rate reach 100%, KFU-B2 and
The Protoplast calli of KAGOU-E2 is respectively 4.0% and 7.5%, shows that breaking yeast cellule membrane is ideal, is conducive to next
Step fusion.
2.2) by fused suspension to plate, the fusant RHZ-2 grown is screened, obtains 16 altogether
Strain fusant bacterial strain, is named as SEB4 ~ 11, F2-1 ~ F2-8.Flocculability has been investigated to above-mentioned 16 plants of fusant bacterial strains, has shown own
Fusant bacterial strain all have the flocculability of the sub- RHZ-1 equal extent of parent fusion.Meanwhile under 30 °C of acid conditions, Suo Yourong
Combined bacteria strain can show that they have been assigned the acid-resistant property of parent SEB2 with glucose fermentation producing and ethanol.
I.e. the present invention is both with the flocculability of the sub- RHZ-1 equal extent of parent fusion, it may have the acidproof spy of parent SEB2
Property.
The present invention carries out fermenting property detection to above-mentioned 16 plants of fusant bacterial strains, is experimentally confirmed: the fermentation of bacterial strain SEB4
Performance is best, is superior to parent strain on ethanol production, glucose utilization and glucose consumption rate.Bacterial strain SEB4's
Specific fermenting property result is as follows:
When 24 h, bacterial strain SEB4 consumes most glucose, residual sugar amount be 31.88 g/l (sugar consumption amount is 78.7%,
Sugar consumption rate is 4.9 g/l/h), ethanol production is 47.87 g/l.
And at this point, the residual sugar amount of parent fusion RHZ-1 and parent SEB2 are respectively that (sugar consumption amount is 52.7 g/l
64.9%, sugar consumption rate is 4.1 g/l/h) and 42.51 g/l (sugar consumption amount is 71.6%, sugar consumption rate be 4.5 g/l/
H), ethanol production is lower than SEB4, respectively 35.93 g/l and 32.69 g/l.
When 48 h, bacterial strain SEB4 and parent strain can all be consumed glucose in 48 h, and the ethyl alcohol of SEB4 produces
Highest is measured, is 57.76 g/l, the ethanol production of parent fusion RHZ-1 and parent SEB2 are respectively 51.93 and 54.42 g/l.
In conclusion this research passes through using the fusant RHZ-1 and SEB2 of IR-2 and SEB1 building as starting strain
The mode of protoplasm somatocyte fusion, final one plant of acquisition have amphilepsis character concurrently, and with outstanding alcohol fermentation performance
Acidproof flocculability Wine brewing yeast strain SEB4 (Lys+, His+)。
Above-described embodiment is merely a preferred embodiment of the present invention, and it is not intended to limit the protection scope of the present invention, as long as using
Design principle of the invention, and the non-creative variation worked and made is carried out on this basis, it should belong to of the invention
Within protection scope.
Claims (1)
1. one plant of acidproof flocculability an industrial strain of S.cerevisiae strain, which is characterized in that the saccharomyces cerevisiae (Saccharomyces
Cerevisiae SEB4) bacterial strain is preserved in China General Microbiological culture presevation administrative center, deposit number CGMCC
No.11324。
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