CN102864178A

CN102864178A - Method for producing pyruvic acid by utilizing torulopsis glabrata

Info

Publication number: CN102864178A
Application number: CN2012103117130A
Authority: CN
Inventors: 金雅新
Original assignee: TAICANG MAOTONG HUAJIAN CO Ltd
Current assignee: TAICANG MAOTONG HUAJIAN CO Ltd
Priority date: 2012-08-29
Filing date: 2012-08-29
Publication date: 2013-01-09

Abstract

The invention provides a method for producing pyruvic acid by utilizing torulopsis glabrata. According to the method, the optimal fermentation culture conditions of bacterial strains are determined by studying the culture conditions of the bacterial strains, the inoculated dose is 10 percent, glucose is 30 g/L, peptone is 10 g/L, corn steep liquor is 10 mL/L, KH2PO4 is 2 g/L, the pH value is 5.6, quantitative vitamins are added, and preferably, the optimum culture temperature for producing the pyruvic acid through torulopsis glabrata is 30 DEG C.

Description

A kind of method of utilizing Torulopsis glabrata to produce pyruvic acid

Technical field

The invention belongs to fermentation technical field, be specifically related to a kind of method of utilizing Torulopsis glabrata to produce pyruvic acid.

Background technology

Pyruvic acid is enzyme process combination colour propylhomoserin and tyrosine, L-3,4 dihydroxyphenylalanine (medically be used for treating Parkinson's disease), being the raw material of hydratropic acid, the initiator of ethylene-based polymer, the synthesis material of artificial organs, is synthetic L-half skin propylhomoserin, L-Leu, VB ₆And VB ₁₂Raw material.Pyruvic acid can also be used to synthesizing series proteinase inhibitor, antiviral agent, tranquilizer, anti-inflammatory analgesic, antiviral agent medicine etc. simultaneously.

Another important applied field of pyruvic acid is exactly as anoretic.The extraneous pyruvic acid that replenishes accelerates the TCA circulation by stimulating plastosome, thereby accelerates the purpose that fat consumption reaches fat-reducing.Mention in the Virtual Musde magazine: " the scientists explanation of the attached university in Pittsburgh medical center: add the supplement that contain pyruvate salt (being generally sodium, potassium, calcium, magnesium salts) in the diet; not only can increase the metabolism of fat up to 48%, and can reduce the protein loss that causes because of low fat diet ".Abroad in Recent Years developed country utilizes this characteristic of pyruvic acid, with its calcium salt as the Weight-reducing health medicine.External relevant media oneself through report, the diet pill that contain CALCIUM PYRUVIC are the fashionable whole world, becomes the new lover on the diet pill market.

Pyruvic Acid Ethyl ester is raw material and the intermediate that is difficult to substitute during agricultural chemicals synthesizes, and it is widely used in the synthetic of sterilant and weedicide.Ethyl bromide acetone is the main intermediate of efficient germicide thiabendazole; thiabendazole is a kind of internal-suction type sterilant of high-efficiency broad spectrum; plant there are protection and therapeutic action; can also control the disease of storage fruit, vegetables; be widely used in the anti-corrosive fresh-keeping of the various fruits vegetables such as oranges and tangerines, banana, China Ministry of Health also permits it and uses as foodstuff additive.

Summary of the invention

The invention provides a kind of method of utilizing Torulopsis glabrata to produce pyruvic acid, by the research to the strain culturing condition, determined the suitableeest fermentation culture conditions of bacterial strain: 10% inoculum size, 30 g/L glucose, 10 g/L peptones, 10 mL/L corn steep liquors, 2 g/L KH ₂PO ₄, pH is 5.6, and adds quantitative VITAMIN, 30 ℃ of cultivations are produced pyruvic acid to Torulopsis glabrata the most favourable.

1. a method of utilizing Torulopsis glabrata to produce pyruvic acid by the research to the strain culturing condition, has been determined the suitableeest fermentation culture conditions of bacterial strain.

2. step 1 the suitableeest described fermentation culture conditions is 10% inoculum size, 30 g/L glucose, 10 g/L peptones, 10 mL/L corn steep liquors, 2 g/L KH ₂PO ₄, pH is 5.6, and adds quantitative VITAMIN, 30 ℃ of cultivations.

3. the described quantitative VITAMIN of step 1 comprises vitamin, nicotinic acid, pyridoxine hydrochloride or vitamin H.

4. the described best vitamin addition of step 1 is 15 μ g/L.

5. the described best nicotinic acid addition of step 1 is 15 mg/L.

6. the described best pyridoxine hydrochloride addition of step 1 is 2 mg/L.

7. the described best vitamin H addition of step 1 is 9 μ g/L.

Beneficial effect of the present invention is: the present invention uses Torulopsis glabrata to produce pyruvic acid, by the research to the strain culturing condition, determines the fermentation culture conditions that bacterial strain is the suitableeest, improves Torulopsis glabrata and produces the pyruvic acid ability.

Description of drawings

Fig. 1 vitamin produces the acetone effect of acid to Torulopsis glabrata.

Fig. 2 nicotinic acid produces the acetone effect of acid to Torulopsis glabrata.

Fig. 3 pyridoxine hydrochloride produces the acetone effect of acid to Torulopsis glabrata.

Fig. 4 vitamin H produces the acetone effect of acid to Torulopsis glabrata.

Embodiment

The following examples elaborate to the present invention, but to not restriction of the present invention.

The used bacterial strain of this patent is Torulopsis glabrata (CCTCC M202019), buys in CCTCC.

Embodiment 1

The implementation case explanation vitamin produces the acetone effect of acid to Torulopsis glabrata, for inquiring into the different concns vitamin Torulopsis glabrata is produced the acetone effect of acid, select the different concns vitamin to carry out pyruvate fermentation, the result as shown in Figure 1.

The results are shown in Figure 1, the thiamine hydrochloride cellulose content is remarkable on the accumulation volume impact of biomass, residual sugar amount and pyruvic acid.Vitamin promotes thalli growth, and when concentration increase along with vitamin concentration during less than 15 μ g/L, biomass increases.After this biomass no longer increases along with the increase of vitamin concentration, basically remains unchanged.This shows, the vitamin of 15 μ g/L is to keep the minimum concentration that thalline is fully grown.Under the condition that lacks vitamin, the decline of PDH enzyme activity causes the pyruvate oxidation decarboxylation to be obstructed, so that pyruvic acid is accumulated.But because vitamin or the cofactor of transketolase, so the phosphopentose pathway synthesis rate that causes NADPH that also is affected reduces, so growing microorganism speed slows down, the pyruvic acid productive rate is higher as a result, but glucose consumption rate is very slow.The explanation of this phenomenon, when vitamin concentration was in the restricted portion (as in the scope less than 15 μ g/L), glycolysis speed also was obstructed.

Embodiment 2

The implementation case explanation nicotinic acid produces the acetone effect of acid to Torulopsis glabrata, for inquiring into different concns nicotinic acid Torulopsis glabrata is produced the acetone effect of acid, selects different concns nicotinic acid to carry out pyruvate fermentation, and the result as shown in Figure 2.

The results are shown in Figure 2, in the scope less than 1 mg/L, increase Nicotinic Acid Content and significantly improve biomass, reduce the residual sugar amount.Continuing to increase Nicotinic Acid Content does not have much impacts to thalli growth, but remaining sugar concentration but constantly descends, and illustrates that nicotinic acid has the ability that promotes thalline to utilize glucose.Nicotinic acid is remarkable not as vitamin on the accumulation impact of pyruvic acid, and the Main Function of inferring nicotinic acid is to accelerate thalline to the speed of utilizing of glucose.And when the nicotinic acid addition was 15 mg/L, output of pyruvic acid was the highest.

Embodiment 3

The implementation case explanation pyridoxine hydrochloride produces the acetone effect of acid to Torulopsis glabrata, for inquiring into the different concns pyridoxine hydrochloride Torulopsis glabrata is produced the acetone effect of acid, select the different concns pyridoxine hydrochloride to carry out pyruvate fermentation, the result as shown in Figure 3.

The results are shown in Figure 3, do not add hydrochloric acid in the fermention medium and adjoin when trembling alcohol, cell concentration is low, the sugared speed of consumption descends, and output of pyruvic acid is low; When hydrochloric acid adjoined the determining alcohol of trembling and reaches 0.5 mg/L, bacterium is dense to increase sharply, and remaining sugar concentration descends rapidly, and output of pyruvic acid increases substantially.And when 2 mg/L, decrease although bacterium is dense, output of pyruvic acid is maximum, and therefore selecting 2 mg/L is optimum addition.

Embodiment 4

The implementation case explanation vitamin H produces the acetone effect of acid to Torulopsis glabrata, for inquiring into the different concns vitamin H Torulopsis glabrata is produced the acetone effect of acid, selects the different concns vitamin H to carry out pyruvate fermentation, and the result as shown in Figure 3.

The results are shown in Figure 4, biotin concentration remarkably influenced thalli growth, pyruvic acid accumulation volume and consume sugared speed in 0-9 μ g/L scope.The vitamin H of lower concentration has reduced pyruvic acid shuttle enzyme activity, causes that growing microorganism speed slows down, glucose consumption rate reduces and the production declining of pyruvic acid.The output of pyruvic acid is the highest when biotin concentration is 9 μ g/L, after this continuing to increase vitamin H content output of pyruvic acid descends on the contrary, illustrate that too much vitamin H has increased the consumption of pyruvic acid, supposition may be that vitamin H has activated pyruvic acid and takes off the shuttle enzyme activity, and the carbon flux that causes pyruvic acid to flow to careless phthalein acetic acid increases.Compare with nicotinic acid with vitamin, when the vitamin H more than the supply 9 μ g/L, its concentration is little on the impact of pyruvate fermentation.

Claims

1. a method of utilizing Torulopsis glabrata to produce pyruvic acid is characterized in that, fermentation culture conditions is 10% inoculum size, 30 g/L glucose, 10 g/L peptones, 10 mL/L corn steep liquors, 2 g/L KH ₂PO ₄, pH is 5.6, and adds quantitative VITAMIN, 30 ℃ of cultivations.

2. method according to claim 1 is characterized in that, quantitatively VITAMIN is selected from vitamin, nicotinic acid, pyridoxine hydrochloride or vitamin H.

3. method according to claim 2 is characterized in that, the vitamin addition is 15 μ g/L.

4. method according to claim 2 is characterized in that, the nicotinic acid addition is 15 mg/L.

5. method according to claim 2 is characterized in that, the pyridoxine hydrochloride addition is 2 mg/L.

6. method according to claim 2 is characterized in that, the vitamin H addition is 9 μ g/L.