CN102174624B - Method for producing enramycin by fermentation - Google Patents

Abstract

The invention discloses a method for producing enramycin by fermentation. Cabicidin streptomyces avermitilis is used as a fermentation strain in the method; and water is supplied to the fermentation solution in the fermentation process of constant temperature culture in a fermentation medium, so that the dissolved oxygen quantity of the fermentation solution is improved, and the fermentation level of the enramycin is improved. The method is simple and feasible; and after a proper amount of water is supplied at proper time in the fermentation culture process, the fermentation level of the enramycin can be improved by about 130 percent.

Description

A kind of method of producing enramycin by fermentation

Technical field

The present invention relates to fermentation technical field, be specially a kind of method of producing enramycin by fermentation.

Background technology

Enramycin (Enramycin), have another name called Enramycin, Enramycin, enramycin, enramycin, it is a kind of ring type polypeptide class microbiotic, by 13 different types of 17 amino acid moleculars and molecular composition of fatty acids, amino acid forms ring type polypeptide, fatty acid molecule is connected on the aspartic acid of end, be divided into enramycin A and enramycin B according to the difference of end fatty acid molecule, enramycin is the mixture of these two kinds of components, is a kind of many skins class microbiotic that actinomycetes (Streptomyces fungieidious) fermentation of separating in soil is produced.Enramycin has very strong restraining effect to gram-positive microorganism, mainly hinders the synthetic of bacteria cell wall.Its sensitive bacterial has various gram-positive coccis, Bacillus subtilus, anthrax bacillus, clostridium tetani, Clostridium botulinum, the clostridium perfringens etc. such as streptococcus aureus.Life-time service enramycin medicine is difficult for developing immunity to drugs, and it can change the community distribution of bacterium in enteron aisle, be conducive to digesting and assimilating of feed nutrition composition, can promote the weight of animals to increase and improve the utilization ratio of feed, therefore microbiotic and the growth promoter of this medicine as livestock and poultry recommended by many countries in the world.Another characteristics of enramycin are to be difficult for being absorbed, so residual less in the livestock and poultry body.There is good growth promotion and improve the effect of efficiency of feed utilization, therefore recommended as the microbiotic growth promoter by many countries in the world.This medicine is in 1966 by day Takede Chemical Industries Ltd research and development, and 1974 at Japanese official registration, is registered and is widely used in many countries thereafter.China Ministry of Agriculture in 1993 ratifies this medicine and registers in China.2005, domestic production enterprise and U.S. Schering Plough animal health-care product company limited (Schering Plough Animal Health Corp.) started the joint production enramycin premix.

Enramycin fermentation thalline is higher to the demand of oxygen, when cell concentration is too high, causes fermenting process oxygen under-supply, affects output.If cell concentration is too low, product enramycin speed is slower, and fermentation period is long, and thalline is aging, does not reach equally desirable effect.Be to solve above contradiction so improve the dissolved oxygen of fermented liquid, improve the effective way of fermentation level.

Summary of the invention

The object of the present invention is to provide a kind of method of producing enramycin by fermentation, in order to the problem of dissolved oxygen deficiency in the fermented liquid that solves existing producing enramycin by fermentation.

For achieving the above object, the method for a kind of producing enramycin by fermentation of the present invention, in the fermenting process of fermentation strain constant temperature culture in fermention medium, toward moisturizing in fermented liquid, improve the dissolved oxygen amount in fermented liquid, improves the fermentation level of enramycin.

The bacterial strain of described fermentation is kabicidin streptomycete (Streptomyces fungicidious) CGMCC 4.1312 ^t.

Described fermention medium comprises following component: Semen Maydis powder 100g/L, corn steep liquor 10g/L, ammonium sulfate 3g/L, zinc sulfate 0.1g/L, calcium carbonate 20g/L.

Wherein, described leavening temperature is 28 ℃, and rotating speed is 250～350r/min.

Describedly enter the quick fermentation phase for fermentation toward the moisturizing time in fermented liquid, preferably fermentation is to during 40～96 hours, more preferably during 40～72 hours, and most preferably 72 hours.

The described rate of water make-up toward moisturizing in fermented liquid is preferably 5%～20% of fermentating liquid volume, and more preferably 5%～10%, most preferably 10%.

Advantage of the present invention:

The invention provides a kind of method of producing enramycin by fermentation, in fermentation, to certain hour, by moisturizing in fermented liquid, improve dissolved oxygen, can significantly improve the enramycin fermentation level.The present invention is simple, in the fermentation culture process after the timely and appropriate discovery moisturizing, the raising of enramycin fermentation level can reach approximately 130%.

Embodiment

For following examples are used for the present invention is described, but be not used for limiting the scope of the invention.

Embodiment 1:

1) slant culture:

Kabicidin streptomycete (Streptomyces fungicidious) CGMCC 4.1312 ^t(purchased from Institute of Micro-biology of the Chinese Academy of Sciences) is inoculated on the solid slant culture base under aseptic condition, 28 ℃ of constant temperature culture 8 days;

Wherein, described solid slant culture base comprises following component: Semen Maydis powder 20g/L, peptone 1g/L, ZnSO ₄.7H ₂o 0.1g/L, MnCl ₂.4H ₂o 0.02g/L, agar 15g/L.

2) fermentation culture:

By step 1) be inoculated on the fermention medium in fermentation flask under the bacterial classification aseptic condition cultivated, 28 ℃, 300r/min, fermentation to 40 hour, to moisturizing in fermented liquid, rate of water make-up is fermentating liquid volume 5%, the fermentation number of days is 8 days, and the average fermentation level is 850U/ml;

Wherein, described fermention medium comprises following component: Semen Maydis powder 100g/L, corn steep liquor 10g/L, ammonium sulfate 3g/L, zinc sulfate 0.1g/L, calcium carbonate 20g/L.

Embodiment 2:

1) slant culture:

Kabicidin streptomycete (Streptomyces fungicidious CGMCC 4.1312 ^t) be inoculated on the solid slant culture base 28 ℃ of constant temperature culture 8 days under aseptic condition;

Wherein, described solid slant culture base comprises following component: Semen Maydis powder 20g/L, peptone 1g/L, ZnSO ₄.7H ₂o 0.1g/L, MnCl ₂.4H ₂o 0.02g/L, agar 15g/L.

2) fermentation culture:

By step 1) be inoculated on the fermention medium in fermentation flask under the bacterial classification aseptic condition cultivated, 28 ℃, 300r/min, fermentation to 72 hour, to moisturizing in fermented liquid, rate of water make-up is fermentating liquid volume 10%, the fermentation number of days is 8 days, the horizontal 1210U/ml of average fermentation;

Wherein, described fermention medium comprises following component: Semen Maydis powder 100g/L, corn steep liquor 10g/L, ammonium sulfate 3g/L, zinc sulfate 0.1g/L, calcium carbonate 20g/L.

Embodiment 3:

1) slant culture:

Kabicidin streptomycete (Streptomyces fungicidious CGMCC 4.1312 ^t) be inoculated on the solid slant culture base 28 ℃ of constant temperature culture 8 days under aseptic condition;

Wherein, described solid slant culture base comprises following component: Semen Maydis powder 20g/L, peptone 1g/L, ZnSO ₄.7H ₂o 0.1g/L, MnCl ₂.4H ₂o 0.02g/L, agar 15g/L.

2) fermentation culture:

By step 1) be inoculated on the fermention medium in fermentation flask under the bacterial classification aseptic condition cultivated, 28 ℃, 300r/min, fermentation to 96 hour, to moisturizing in fermented liquid, rate of water make-up is fermentating liquid volume 20%, the fermentation number of days is 8 days, the horizontal 960U/ml of average fermentation;

Wherein, described fermention medium comprises following component: Semen Maydis powder 100g/L, corn steep liquor 10g/L, ammonium sulfate 3g/L, zinc sulfate 0.1g/L, calcium carbonate 20g/L.

Comparative example:

1) slant culture:

Kabicidin streptomycete (Streptomyces fungicidious CGMCC 4.1312 ^t) be inoculated on the solid slant culture base 28 ℃ of constant temperature culture 8 days under aseptic condition;

Wherein, described solid slant culture base comprises following component: Semen Maydis powder 20g/L, peptone 1g/L, ZnSO ₄.7H ₂o 0.1g/L, MnCl ₂.4H ₂o 0.02g/L, agar 15g/L.

2) fermentation culture:

By step 1) be inoculated on the fermention medium in fermentation flask under the bacterial classification aseptic condition cultivated, 28 ℃, 300r/min, fermented.The fermentation number of days is 8 days; The horizontal 530U/ml of average fermentation;

Wherein, described fermention medium comprises following component: Semen Maydis powder 100g/L, corn steep liquor 10g/L, ammonium sulfate 3g/L, zinc sulfate 0.1g/L, calcium carbonate 20g/L.

Above embodiment is used for the present invention is described, but is not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, modification made for the present invention or replacement, all belong to scope of the present invention.

Claims (3)

1. the method for a producing enramycin by fermentation, is characterized in that, in the fermenting process of fermentation strain constant temperature culture in fermention medium, toward moisturizing in fermented liquid, improves the dissolved oxygen amount in fermented liquid, improves the fermentation level of enramycin; Wherein, described leavening temperature is 28 ℃, and rotating speed is 300r/min;

The described moisturizing time is fermentation 40～96 hours;

Toward the amount of moisturizing in fermented liquid, be fermentating liquid volume 5%～20%;

Bacterial strain for producing enramycin by fermentation is kabicidin streptomycete (Streptomyces fungicidious) CGMCC4.1312 ^t;

Described fermention medium comprises following component: Semen Maydis powder 100g/L, corn steep liquor 10g/L, ammonium sulfate 3g/L, zinc sulfate 0.1g/L, calcium carbonate 20g/L.

2. method according to claim 1, is characterized in that, the described moisturizing time is fermentation 40～72 hours.

3. method according to claim 1, is characterized in that, toward the amount of moisturizing in fermented liquid, is fermentating liquid volume 5%～10%.