CN103642707B - Method for producing candida for feeds by fermenting high-salt-content amino acid wastewater - Google Patents

Method for producing candida for feeds by fermenting high-salt-content amino acid wastewater Download PDF

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CN103642707B
CN103642707B CN201310643599.6A CN201310643599A CN103642707B CN 103642707 B CN103642707 B CN 103642707B CN 201310643599 A CN201310643599 A CN 201310643599A CN 103642707 B CN103642707 B CN 103642707B
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amino acid
acid wastewater
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high saliferous
candida
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CN103642707A (en
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胡征
杨波
刘志刚
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Hubei University of Technology
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Abstract

The invention discloses a method for producing candida for feeds by fermenting high-salt-content amino acid wastewater, which comprises the following steps: 1. habituated culture of feed strain: carrying out continuous habituated culture to enhance the tolerance of the strain to high-salt-content amino acid wastewater; and 2. fermentation of strain: under appropriate fermenting conditions, carrying out liquid ventilation culture in a fermentation tank for some time to obtain the microzyme for feeds. The microzyme is centrifugated, collected, dried in a 70 DEG C drying oven to constant weight, and weighed. The calculation indicates that every liter of high-salt-content amino acid wastewater can obtain 145g of candida at the maximum. The method can effectively utilize amino acids, microelements and growth factors contained in the wastewater, greatly lowers the production cost of the candida for feeds, greatly reduces the sewage treatment load, achieves the comprehensive utilization effect, and has obvious economic and social benefits.

Description

Utilize the method for the feeding candiyeast of high saliferous amino acid wastewater fermentative production
Technical field
The present invention relates to the field of comprehensive utilization of waste water resource, is a kind ofly relate to the method utilizing high saliferous amino acid wastewater fermentation production of fodder candiyeast in particular.
Background technology
According to World Health Organization's investigation, there is the people of 70% to drink tap water less than safety and sanitation in the world, have every day 2.5 ten thousand people to die from the disease relevant to sewage or severe sanitary condition; To the year two thousand fifty, estimate that the whole world has about 7,000,000,000 people and faces and use water shortage.China has the people of 82% to drink shallow well and river, and what wherein bacterial contamination exceeded hygienic standard accounts for 75%, is subject to the drinking-water population about 1.6 hundred million of Organic pollutants.Wherein the sewage of industrial discharge is the main source of water pollutions.Through investigation statistics, the annual wastewater discharge 659.2 hundred million tons in the whole nation.Wherein, discharged volume of industrial waste water 230.9 hundred million tons, accounts for 35.0% of wastewater emission amount.In fact, water amount of blowdown is considerably beyond this number, because many township and village enterprises industrial wastewater discharge amount is difficult to statistics.
China is amino acids production big country, the production method that the enterprise that major part produces halfcystine takes is all hair Acid hydrolysis method, the high saliferous amino acid wastewater not only enormous amount of annual generation, and Pollutant levels are high, its COD concentration is 50 ~ 80g/L, ammonia nitrogen concentration is 5 ~ 8g/L, total cl concn is 80-180g/L, belong to that typical high density is organic, ammonia nitrogen, high salinity, strongly acid wastewater, greatly, the problem of environmental pollution caused thus has become the key of restriction amino acids production industry value chain for intractability and burden.But, high-concentration waste water in amino acids production is containing there being again a lot of valuable resource, as the solid matter such as amino acid, protein suspended substance, plurality of inorganic salt, organic acid and reducing sugar etc., full nitrogen (N) content is up to 75-150g/L, and aminoacids content is up to 150-300g/L.Do not contain the difficult poisonous and harmful substances decomposed in this waste water simultaneously.This large amount of discharges containing amino acid wastewater not only pollute environment and also waste amino acid much with practical value.Under current international economy amplification turns slow overall background, attempting " energy-saving and emission-reduction " cleaner production theory and bring new opportunity by for amino acids production enterprise, is also the great scientific research task that fermentation industry faces.
At present, for the process of high saliferous amino acid wastewater, there is no both at home and abroad ripe can the treatment process of popularity.The amino acid wastewater treatment process reported at present mainly contains electroosmose process, ultrafiltration process, biochemical process etc., but electrodialysis and ultrafiltration process are because of investment with running cost is high, safeguard that the shortcoming such as inconvenient limits the popularization of the method; And biochemical process because of its investment and running cost low, technology maturation, does not produce secondary pollution, operational management facilitate etc. advantage in treatment of Organic Wastewater, obtain global concern.Current this method Problems existing is that centering sewage of low concentration is comparatively suitable, but is not suitable for the amino acid wastewater processing high density.The amino acid wastewater of high density is because of higher concentration NH 3the salt of-N and high density, can produce strong restraining effect to anaerobism and aerobic microorganism, make biochemical reaction effect poor, the not easily stably reaching standard discharge of the water after process.If by sewage dilution for many times, considerably increase treatment capacity again, the quality amino acids resource simultaneously in waste water has all wasted.At present also someone adopts and first carries out pre-treatment (its aminoacid component of concentration and recovery) to high-purity amino acid waste water, and then with the composite waste of biotechnology process lower concentration.This method is a kind of effective ways of process amino acid wastewater, has both reclaimed amino acid, and has again reduced the burden of biological treatment.But it is high whole treatment process to exist concentration process cost, time and effort consuming, and resource reclaim disconnects mutually with wastewater treatment, and the shortcoming such as after having high temperature evaporation concentrated organism is apt to deteriorate.
Summary of the invention
The object of the invention is to be to provide a kind of method utilizing high saliferous amino acid wastewater fermentation production of fodder candiyeast, its technique is simple, with low cost, the process of high saliferous amino acid wastewater and fermentative production are directly combined, eliminate all middle pre-treatment step, realize the direct biological utilisation of high-concentration sewage, while realizing at low cost turning waste into wealth, also reduce discharge, economic benefit and social benefit remarkable.
In order to realize above-mentioned object, the present invention adopts following technical measures:
Utilize a method for high saliferous amino acid wastewater fermentation production of fodder candiyeast, the steps include:
(1) continuous domestication of feed bacterial strain is cultivated, specific as follows: first preparation is containing the liquid nutrient medium of high saliferous amino acid sewage of different concns gradient, adds 1.5%-2.5%(mass volume ratio) agar makes solid medium.After feed candiyeast is activated according to a conventional method, the solid medium being inoculated into above-mentioned different concns gradient successively carries out continuous passage cultivation (culture temperature is 15-35 DEG C), progressively improves the salt resistance ability of bacterial classification.Through bimestrial cultured continuously, final bacterial strain can account in the above-mentioned substratum of substratum about total mass 45-55% at high saliferous amino acid wastewater and grow.
(2) ferment: the candida bacterial strain of step (1) being tamed is seeded to in the high saliferous amino acid wastewater fermention medium that is nitrogenous source by inoculum size 0.5-5%, at 15-35 DEG C, rotating speed is in fermentor tank, carry out liquid fermenting under the condition of 100-300rpm/min, ventilation coefficient is that 0.2-0.8(per minute passes into fermentating liquid volume 0.2-0.8 filtrated air doubly), be placed in 70 DEG C of thermostatic drying chambers after the centrifugal 10min of 4000r/min collects thalline after fermentation 2-7d, weigh after being dried to constant weight.As calculated, according to said method often raise saliferous amino acid wastewater and can obtain 145g yeast thalline through bio-transformation is the highest.
Described high saliferous amino acid wastewater produces the waste water produced in amino acid whose enterprise industrial production from hair Acid hydrolysis, full nitrogen (N) content is 75-150g/L, and aminoacids content is 150-300g/L, and chloride ion content is 80-180g/L.
Described feed candiyeast is Candida utilis (Candida utilis) CICC1769 or candida tropicalis (Candida tropicalis) CICC1419 that derive from Chinese industrial Microbiological Culture Collection administrative center;
The liquid nutrient medium of the described high saliferous amino acid wastewater containing different concns gradient, the concentration of its high saliferous amino acid wastewater is respectively 125g/L, 250g/L, 330g/L, 400g/L and 500g/L, other chemical matrix components are: carbon source (by glucose meter): 30-140g/L, potassium primary phosphate (KH 2pO 4): 5-22.5g/L, yeast powder: 3-15g/L, magnesium sulfate (MgSO 4): 3-13.5g/L, calcium carbonate (CaCO 3): 0.75-3g/L, ferrous sulfate (FeSO 4): 1.5-6g/L, medium pH is 3.5-7.5; Wherein, carbon source is the mixture of one or more following carbohydrates: glucose; Sucrose; Molasses; W-Gum enzymolysis solution; Sweet potato starch enzymolysis solution or tapioca (flour) enzymolysis solution.
Described W-Gum enzymolysis solution; Sweet potato starch enzymolysis solution or tapioca (flour) enzymolysis solution be with standard enzymatic hydrolysis be prepared from (or see Liu Yawei. production of corn starch and transformation technology [M]. Beijing: Chemical Industry Press, 2003:296.)
The described fermention medium being nitrogenous source with high saliferous amino acid wastewater, its formula is: high saliferous amino acid sewage: 125g/L-330g/L, carbon source: 30-100g/L, potassium primary phosphate (KH 2pO 4): 5-15g/L, yeast powder: 0-10g/L, magnesium sulfate (MgSO 4): 3-9g/L, calcium carbonate (CaCO 3): 0.75-2g/L, ferrous sulfate (FeSO 4): 1.5-4g/L, medium pH is 3.5-7.5; Wherein, carbon source is the mixture of one or more following carbohydrates: glucose, sucrose, molasses, W-Gum enzymolysis solution, sweet potato starch enzymolysis solution or tapioca (flour) enzymolysis solution.
As preferably, the culture temperature that in step (1), domestication is cultivated is 20-35 DEG C, and the medium pH that domestication is cultivated is 5-6.5.
As preferably, the fermention medium described in step (2), medium pH is 5-6.5.
As preferably, the culture temperature in step (2) is 20-35 DEG C, and incubation time is 3-5d.
Compared with prior art, the present invention has the following advantages:
Bacterial strain in the present invention is after continuous domestication is cultivated, can grow fast using high density height saliferous amino acid wastewater as nitrogenous source, so just successfully the purifying treatment of high saliferous amino acid wastewater and fermentative production are directly combined, eliminate middle pre-treatment step, achieve the low cost recycling of amino acid wastewater, both can reclaim a large amount of useful matter, reduce the cost of fermentative production, also reduce the discharge of sewage simultaneously, there is very strong competitive edge.For Wuhan halfcystine manufacturing enterprise, it is with various animal hair for each seed amino acid of raw material production, and year produces waste liquid about 10,000 tons; As waste liquid utilizes completely, can year fermentative production 1200-1500 ton fodder yeast, reach the annual value of production of 6000-7500 ten thousand, can also reduce the discharge of high cod values waste water, alleviate waste disposal plant load, Benefits in economic and social terms is remarkable simultaneously.
The salt-enduring strain obtained in the present invention carries out domestication with traditional feed candida bacterial strain for starting strain traditional method to cultivate and create physiological compatibility and obtain, do not change its legacy feature, there is extraordinary biological safety, can production be directly used in.
The salt-enduring strain vitality of gained of the present invention is strong, and nitrogenous source utilising efficiency is high, under fermentor tank bench scale, can reach the production level of often liter of waste water stoste output 145g yeast thalline, and single just have very high application potentiality economically.
The present invention is utilized to tame the bacterial strain of cultivation, the process of high saliferous amino acid wastewater and fermentative production directly can be combined, eliminate all middle pre-treatment step, realize the direct biological utilisation of high-concentration sewage, while realizing at low cost turning waste into wealth, also reduce discharge, economic benefit and social benefit remarkable.
Embodiment
Below in conjunction with specific embodiment, the present invention is further elaborated; But the present invention is not limited to these embodiments.
The microorganism used in following examples is all the conventional bacterial classification that the public can obtain.Obviously, those skilled in the art also can use other candida strain to verify technical scheme of the present invention.Described technical scheme, if not otherwise specified, is conventional techniques, and agents useful for same is this area common agents.
Embodiment 1: the domestication of feed candida strain is cultivated.
1. fetch from Wuhan Amino Acid Factory, hair Acid hydrolysis produces the waste liquid after halfcystine, as the use of high saliferous amino acid wastewater required below.Its component list is as follows:
Table 1 amino acid wastewater coherent element content
Project name Unit Detected result Method foundation
Full nitrogen (N) 8.79 GB/T8572-2010
Sodium (Na) 0.52 ICP method
Chlorine (Cl) g/L 131.26 Ag Cl turbidimetry
Plumbous (Pb) mg/kg 0.91 ICP method
Arsenic (As) mg/kg 0.076 Atomic fluorescence method
Mercury (Hg) mg/kg 0.014 Atomic fluorescence method
Chromium (Cr) mg/kg 1.51 ICP method
Cadmium (Cd) mg/kg 0.06 ICP method
Table 2 amino acid wastewater aminoacid component and content
Project name Detected result %
Taurine 0.02
Aspartic acid 2.16
Threonine 1.58
Serine 3.92
L-glutamic acid 3.33
Glycine 3.18
L-Ala 1.58
Gelucystine 0.19
α-amino-isovaleric acid 1.50
Methionine(Met) 0.10
Isoleucine 0.56
Leucine 0.73
Tyrosine 0.16
Phenylalanine 1.20
Γ-aminobutyric acid Do not detect
Ornithine 0.03
Methionin 0.42
Proline(Pro) 2.99
Histidine 0.18
Tryptophane Do not detect
Arginine 2.07
Oxyproline Do not detect
Summation 25.9
Learn from upper table, its amino acid total mass mark reaches 259g/L, and total chlorine, up to 131.26g/L, is typical high saliferous amino acid wastewater.
2. preparation is containing the liquid nutrient medium of high saliferous amino acid wastewater.Preparing high saliferous amino acid wastewater concentration is respectively 125g/L, 250g/L, 330g/L, 400g/L and 500g/L(high saliferous amino acid wastewater of the present invention is in mass) liquid nutrient medium of different gradient, wherein amino acid wastewater is only nitrogen source, described liquid nutrient medium also adds other chemical matrix needed for microorganism, and its component is: glucose: 90g/L, KH 2pO 4: 15g/L, yeast powder: 10g/L, MgSO 4: 9g/L, CaCO 3: 2g/L, FeSO 4: 4g/L, medium pH is 5.5.And change into solid medium after partially liq substratum being added interpolation 20g/L agar (calculating by 1000ml water).
3. feed Candida utilis (Candida utilis) and candida tropicalis (Candida tropicalis) are accessed according to a conventional method respectively after YEPD substratum activates, the concentration be inoculated into successively containing amino acid wastewater is respectively 125g/L, 250g/L, 330g/L, the amino acid wastewater solid medium of 400g/L and 500g/L carries out continuous passage cultivation (culture temperature is 28 DEG C), reach the object that domestication is cultivated.Cultivated by domestication, above-mentioned two bacterial strains can be all the cultured on solid medium of 500g/L in high saliferous amino acid wastewater concentration.
Described Candida utilis (Candida utilis) and candida tropicalis (Candida tropicalis) are bought in Chinese industrial Microbiological Culture Collection administrative center, and its deposit number is respectively CICC1769 and CICC1419.
Embodiment 2:
Candida utilis (Candida utilis) take glucose as carbon source, fermentation in the different concns amino acid wastewater liquid nutrient medium that is nitrogenous source.
1. prepare the liquid fermentation medium of two kinds of different concns height saliferous amino acid wastewaters.Preparing high saliferous amino acid wastewater concentration is respectively the liquid fermentation medium of 165g/L and 330g/L, wherein high saliferous amino acid wastewater is only nitrogen source (its source, composition consistent with embodiment 1), described liquid fermentation medium also adds other chemical matrix needed for microorganism, its component is respectively: glucose: 45 and 90g/L, KH 2pO 4: 7.5 and 15g/L, yeast powder: 5 and 10g/L, MgSO 4: 4.5 and 9g/L, CaCO 3: 1 and 2g/L, FeSO 4: 2 and 4g/L, medium pH is 5.5.Being compiled respectively by this two substratum is No. 1 and No. 2.
2. by Candida utilis (Candida utilis) bacterial strain of having tamed obtained from embodiment 1 by 2% inoculum size be seeded to step 1 prepare No. 1 and No. 2 two kinds of liquid fermentation mediums in, at 28 DEG C in fermentor tank, rotating speed is 200rpm/min, and ventilation coefficient is that under the condition of 0.5, liquid ventilation cultivates 3d.Cultivation terminates the centrifugal 10min of rear 4000r/min and collects thalline respectively, is placed in 70 DEG C of thermostatic drying chambers, dry until weigh after constant weight.Found that, after converting, adopt No. 1 fermentation that substratum carries out, often liter of waste water stoste output 87g yeast thalline; Adopt No. 2 fermentations that substratum carries out, often liter of waste water stoste output 110g yeast thalline.
No. 1 described substratum is specially: high saliferous amino acid wastewater 165g/L, glucose 45g/L, KH 2pO 47.5g/L, yeast powder 5g/L, MgSO 44.5g/L, CaCO 31g/L, FeSO 42g/L, medium pH 5.5;
No. 2 described substratum are specially: high saliferous amino acid wastewater 330g/L, glucose 90g/L, KH 2pO 415g/L, yeast powder 10g/L, MgSO 49g/L, CaCO 32g/L, FeSO 44g/L, medium pH 5.5.
Embodiment 3:
Candida utilis (Candida utilis) be carbon source with W-Gum enzymolysis solution, fermentation in the different concns amino acid wastewater liquid nutrient medium that is nitrogenous source.
1. prepare the liquid fermentation medium of two kinds of different concns height saliferous amino acid wastewaters.Preparing high saliferous amino acid wastewater concentration is respectively the liquid fermentation medium of 165g/L and 330g/L, wherein high saliferous amino acid wastewater is only nitrogen source (its source, composition consistent with embodiment 1), described liquid fermentation medium also adds other chemical matrix needed for microorganism, its component is respectively: W-Gum enzymolysis solution (according to W-Gum dry weight note): 55 and 110g/L, KH 2pO 4: 7.5 and 15g/L, yeast powder 0 and 10g/L, MgSO 4: 4.5 and 9g/L, CaCO 3: 1 and 2g/L, FeSO 4: 2 and 4g/L, medium pH is 5.5.Being compiled respectively by this two substratum is No. 1 and No. 2.
2. Candida utilis (Candida utilis) bacterial strain of having tamed obtained from embodiment 1 is seeded to by 5% inoculum size in two kinds of liquid fermentation mediums of step 1 preparation, at 28 DEG C in fermentor tank, rotating speed is 200rpm/min, and ventilation coefficient is that under the condition of 0.5, liquid ventilation cultivates 3d and 5d.Cultivation terminates the centrifugal 10min of rear 4000r/min and collects thalline respectively, is placed in 70 DEG C of thermostatic drying chambers, dry until weigh after constant weight.Found that, after converting, adopt No. 1 fermentation that substratum carries out, often liter of waste water stoste output 120g yeast thalline, adopt the fermentation that No. 2 substratum carry out, often liter of waste water stoste output 145g yeast thalline.
No. 1 described substratum is specially: high saliferous amino acid wastewater 165g/L, W-Gum enzymolysis solution 55g/L, KH 2pO 4: 7.5g/L, MgSO 44.5g/L, CaCO 31g/L, FeSO 42g/L, medium pH 5.5.
No. 2 described substratum are specially: high saliferous amino acid wastewater 330g/L, W-Gum enzymolysis solution 110g/L, KH 2pO 415g/L, yeast powder 10g/L, MgSO 49g/L, CaCO 32g/L, FeSO 44g/L, medium pH 5.5.
Described W-Gum enzymolysis solution, the method for enzymatic hydrolysis is see Liu Yawei. production of corn starch and transformation technology [M]. and Beijing: Chemical Industry Press, 2003:296.
Embodiment 4:
Candida tropicalis (Candida tropicalis) take glucose as carbon source, fermentation in the different concns amino acid wastewater liquid nutrient medium that is nitrogenous source.
1. prepare the liquid fermentation medium of two kinds of different concns height saliferous amino acid wastewaters.Preparing high saliferous amino acid wastewater concentration is respectively the liquid fermentation medium of 165g/L and 330g/L, wherein high saliferous amino acid wastewater is only nitrogen source (its source, composition consistent with embodiment 1), described liquid fermentation medium also adds other chemical matrix needed for microorganism, its component is respectively: glucose: 45 and 90g/L, KH 2pO 4: 7.5 and 15g/L, yeast powder: 5 and 10g/L, MgSO 4: 4.5 and 9g/L, CaCO 3: 1 and 2g/L, FeSO 4: 2 and 4g/L, medium pH is 5.5.Being compiled respectively by these two kinds of substratum is No. 1 and No. 2.
2. by candida tropicalis (Candida tropicalis) bacterial strain of having tamed obtained from embodiment 1 by 1% inoculum size be seeded to step 1 prepare No. 1 and No. 2 two kinds of liquid fermentation mediums in, at 28 DEG C in fermentor tank, rotating speed is 200rpm/min, and ventilation coefficient is liquid fermentation and culture 3d under the condition of 0.5.Cultivation terminates the centrifugal 10min of rear 4000r/min and collects thalline respectively, is placed in 70 DEG C of thermostatic drying chambers, dry until weigh after constant weight.Found that, after converting, adopt No. 1 fermentation that substratum carries out, often liter of waste water stoste output 82g yeast thalline; Adopt No. 2 fermentations that substratum carries out, often liter of waste water stoste output 105g yeast thalline.
No. 1 described substratum is specially: high saliferous amino acid wastewater 165g/L, glucose 45g/L, KH 2pO 47.5g/L, yeast powder 5g/L, MgSO 44.5g/L, CaCO 31g/L, FeSO 42g/L, medium pH 5.5;
No. 2 described substratum are specially: high saliferous amino acid wastewater 330g/L, glucose 90g/L, KH 2pO 415g/L, yeast powder 10g/L, MgSO 49g/L, CaCO 32g/L, FeSO 44g/L, medium pH 5.5.
Embodiment 5:
Candida tropicalis (Candida tropicalis) be carbon source with W-Gum enzymolysis solution, fermentation in the different concns amino acid wastewater liquid nutrient medium that is nitrogenous source.
1. prepare the liquid fermentation medium of two kinds of different concns height saliferous amino acid wastewaters.Preparing high saliferous amino acid wastewater concentration is respectively the liquid fermentation medium of 165g/L and 330g/L, wherein high saliferous amino acid wastewater is only nitrogen source (its source, composition consistent with embodiment 1), described liquid fermentation medium also adds other chemical matrix needed for microorganism, its component is respectively: W-Gum enzymolysis solution (according to W-Gum dry weight note): 55 and 110g/L, KH 2pO 4: 7.5 and 15g/L, yeast powder 0 and 10g/L, MgSO 4: 4.5 and 9g/L, CaCO 3: 1 and 2g/L, FeSO 4: 2 and 4g/L, medium pH is 5.5.Being compiled respectively by this two substratum is No. 1 and No. 2.
2. candida tropicalis (Candida tropicalis) bacterial strain of having tamed obtained from embodiment 1 is seeded to by 5% inoculum size in two kinds of liquid fermentation mediums of step 1 preparation, at 28 DEG C in fermentor tank, rotating speed is 200rpm/min, and ventilation coefficient is that under the condition of 0.5, liquid ventilation cultivates 3d and 5d.Cultivation terminates the centrifugal 10min of rear 4000r/min and collects thalline respectively, is placed in 70 DEG C of thermostatic drying chambers, dry until weigh after constant weight.Found that, after converting, adopt No. 1 fermentation that substratum carries out, often liter of waste water stoste output 115g yeast thalline, adopt the fermentation that No. 2 substratum carry out, often liter of waste water stoste output 139g yeast thalline.
No. 1 described substratum is specially: high saliferous amino acid wastewater 165g/L, W-Gum enzymolysis solution 55g/L, KH 2pO 4: 7.5g/L, MgSO 44.5g/L, CaCO 31g/L, FeSO 42g/L, medium pH 5.5.
No. 2 described substratum are specially: high saliferous amino acid wastewater 330g/L, W-Gum enzymolysis solution 110g/L, KH 2pO 415g/L, yeast powder 10g/L, MgSO 49g/L, CaCO 32g/L, FeSO 44g/L, medium pH 5.5.
Described W-Gum enzymolysis solution, the method for enzymatic hydrolysis is see Liu Yawei. production of corn starch and transformation technology [M]. and Beijing: Chemical Industry Press, 2003:296.
Although made detailed description to the present invention and quoted some specific embodiments as proof, to those skilled in the art, only otherwise it is obvious for leaving that the spirit and scope of the present invention can make various changes or revise.

Claims (4)

1. utilize a method for high saliferous amino acid wastewater fermentation production of fodder candiyeast, the steps include:
(1) continuous domestication of feed bacterial strain is cultivated, specific as follows: first preparation is containing the liquid nutrient medium of the high saliferous amino acid wastewater of different concns gradient, and the agar adding 1.5%-2.5% mass volume ratio makes solid medium; After being activated according to a conventional method by feed candiyeast, the solid medium being inoculated into above-mentioned different concns gradient successively carries out continuous passage cultivation, and culture temperature is 15-35 DEG C, progressively improves the salt resistance ability of bacterial classification; Final bacterial strain can account in the above-mentioned substratum of substratum total mass 45-55% at high saliferous amino acid wastewater and grow;
(2) ferment: the candida bacterial strain of step (1) being tamed is seeded to in the high saliferous amino acid wastewater fermention medium that is nitrogenous source by inoculum size 0.5-5%, at 15-35 DEG C, rotating speed is in fermentor tank, carry out liquid fermenting under the condition of 100-300rpm, ventilation coefficient is 0.2-0.8, be placed in 70 DEG C of thermostatic drying chambers after the centrifugal 10min of 4000r/min collects thalline after fermentation 2-7d, weigh after being dried to constant weight;
Described high saliferous amino acid wastewater produces the waste water produced in amino acid whose enterprise industrial production from hair Acid hydrolysis, total nitrogen content is 75-150g/L, and aminoacids content is 150-300g/L, and chloride ion content is 80-180g/L;
Described feed candiyeast is Candida utilis (Candida utilis) CICC1769 or candida tropicalis (Candida tropicalis) CICC1419 that derive from Chinese industrial Microbiological Culture Collection administrative center;
The liquid nutrient medium of the described high saliferous amino acid wastewater containing different concns gradient, the concentration of its high saliferous amino acid wastewater is respectively 125g/L, 250g/L, 330g/L, 400g/L and 500g/L, other chemical matrix components are: carbon source: 30-140g/L, potassium primary phosphate: 5-22.5g/L, yeast powder: 3-15g/L, magnesium sulfate: 3-13.5g/L, calcium carbonate: 0.75-3g/L, ferrous sulfate: 1.5-6g/L, medium pH is 3.5-7.5; Wherein, carbon source is the mixture of one or more following carbohydrates: glucose, sucrose, molasses, W-Gum enzymolysis solution, sweet potato starch enzymolysis solution or tapioca (flour) enzymolysis solution;
The described fermention medium being nitrogenous source with high saliferous amino acid wastewater, its formula is: high saliferous amino acid sewage: 125g/L-330g/L, carbon source: 30-100g/L, potassium primary phosphate: 5-15g/L, yeast powder: 0-10g/L, magnesium sulfate: 3-9g/L, calcium carbonate: 0.75-2g/L, ferrous sulfate: 1.5-4g/L, medium pH is 3.5-7.5; Wherein, carbon source is the mixture of one or more following carbohydrates: glucose, sucrose, molasses, W-Gum enzymolysis solution, sweet potato starch enzymolysis solution or tapioca (flour) enzymolysis solution.
2. method according to claim 1, is characterized in that: the liquid nutrient medium pH of the high saliferous amino acid wastewater containing different concns gradient described in described step (1) is 5-6.5, and culture temperature is 20-35 DEG C.
3. method according to claim 1, is characterized in that: the fermention medium described in described step (2), and medium pH is 5-6.5.
4. method according to claim 1, is characterized in that: the culture temperature of described step (2) is 20-35 DEG C, and incubation time is 3-5d.
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