CN102660598B - Method for improving fermentation yield of rifamycin SV - Google Patents

Abstract

The invention discloses a method for improving the fermentation yield of rifamycin SV. In the process of fermentation culture, oxalate with the final concentration of 0.2-0.5g/L and soybean oil with the final concentration of 2-5mL/L are sterilely added into a fermentation tank the fermentation culture cycle of amycolatopsis mediterranei of which is 60-80 hours. The titer of the rifamycin SV is increased by 3.0% on average, the titer is increased by 5.4% on average in case of one billion tanks, carbohydrate supplementation is decreased by 17% and the raw material cost of every tank is saved by***. The supplementary materials in the method are all agricultural and industrial commodities with high affordability. In addition, the operational process is simple and the controllability is strong, thus the method is applicable to large-scale fermentation production, the production increase effect is stable and the method conforms to the green development requirement of energy conservation in fermentation industry by the state.

Description

A kind of method that improves Rifamycin Sodium fermentation yield

Technical field

The present invention relates to a kind of preparation method of rifomycin, particularly a kind of method that improves Rifamycin Sodium fermentation yield.

Background technology

As large microbiotic bulk drug of tuberculotherapy, domestic Rifamycin Sodium fermentation unit is hovered in 5000 U/mL left and right always.The large production level thalline physiological status of industry and technology controlling and process are closely related, the synthetic precursor route of synthesis carbon stream of glycolysis-, phosphopentose pathway, krebs cycle pathway and rifomycin exists makes effect mutually, determine that the latter and the corresponding relation of concrete metabolism bottleneck will make to instruct Rifamycin Sodium fermentation manufacturing technique to optimize indexing, specialize, and have important practical significance for the handling and implementation capacity of enhancing productivity.

The carbon nitrogen metabolism approach of microorganism is interlaced with one another and becomes metabolism network, and carbon, nitrogen stream are determining the resultant quantity of target product in the allocation proportion of metabolism node.For flexible metabolism node, regulate and control this node metabolism stream distribute the structural adjustment that may cause whole metabolism network metabolism stream to distribute (such as: by strengthening precursor anabolism stream or reducing bypass metabolism and flow), thereby likely make carbon nitrogen source metabolism with best proportional flow to target product, and the change of macro-scale operational variable can produce across the impact of yardstick the optimization of bacterium micro-scale physiologic variables.Therefore, by the adjustment of carbon metabolism flow allocation proportion between glycolytic pathway, phosphopentose pathway, shikimic acid pathway and tricarboxylic acid cycle, can promote the synthetic amount that supplies of three larger precursors, promote rifomycin synthetic.

At present, more to the research of raising rifomycin biosynthesizing efficiency both at home and abroad, comprise: the aspects such as strain improvement, process optimization, cheap starting material replacement, yet, through science and technology, look into newly, relevant the oxalate of optimal concentration, soya-bean oil are filled in Amycolatopsis mediterranei fermenting culture at optimum time together, regulation and control Rifamycin Sodium precursor anabolism stream, the research and the application that improve its combined coefficient yet there are no pertinent literature and publication report.

Summary of the invention

The object of the present invention is to provide a kind of method that improves Rifamycin Sodium fermentation yield.

The technical solution adopted in the present invention is as follows:

(1) fermentation culture

Press seed and fermentative medium formula configuration nutrient solution, after moist heat sterilization, be cooled to 28 ℃, by the inoculum size of 3%-5%, bottle seed liquor of closing of Amycolatopsis mediterranei is accessed to a first class seed pot.First class seed pot temperature control is at 28 ℃, and sterile air flow is controlled by 0.8 vvm, and mixing speed is 160 r/min, and tank pressure is controlled at 0.03-0.04 MPa, and after cultivation 48h, aseptic technique fills into secondary seed tank; The temperature control of secondary seed tank is at 28 ℃, and sterile air flow is controlled by 0.8 vvm, and mixing speed is 160 r/min, and tank pressure is controlled at 0.03-0.04 MPa, and after cultivation 48h, aseptic technique fills into three grade fermemtation tank; Three grade fermemtation tank temperature is by 28 ℃ of controls, and rotating speed is 160 r/min, and sterile air flow control is at 0.5-0.8 vvm, and tank pressure is controlled at 0.03-0.05 MPa, and culture cycle is 136 h;

(2) supplying technics

Three grade fermemtation culture cycle is when 60-80 h, and by final concentration, being that the oxalate of 0.2-0.5 g/L and the soya-bean oil of 2-5 mL/L are aseptic fills in three grade fermemtation tank, can improve Rifamycin Sodium output.

Beneficial effect of the present invention is:

1) the feed supplement material using is industrial or agricultural bulk product, and affordability is strong;

2) operating procedure using is simple, and controllability is strong, is suitable for fermentative production in enormous quantities;

3) utilize this feed supplement method, effect of increasing production is stablized, and Rifamycin Sodium is tired and on average improved 3.0%, puts the average raising 5.4% of tank 1,000,000,000, has reduced by 17% sugared the measuring of benefit, and every tank material cost is saved 360 yuan;

4) this feed supplement method meets country about the energy-saving and cost-reducing Green Development requirement of fermentation industry.

Embodiment

Embodiment 1

1 fermentation culture

Press seed and fermentative medium formula configuration nutrient solution, after moist heat sterilization, be cooled to 28 ℃, by the inoculum size of 3%-5%, bottle seed liquor of closing of Amycolatopsis mediterranei is accessed to a first class seed pot.First class seed pot temperature control is at 28 ℃, and sterile air flow is controlled by 0.8 vvm, and mixing speed is 160 r/min, and tank pressure is controlled at 0.03-0.04 MPa, and after cultivation 48h, aseptic technique fills into secondary seed tank; The temperature control of secondary seed tank is at 28 ℃, and sterile air flow is controlled by 0.8 vvm, and mixing speed is 160 r/min, and tank pressure is controlled at 0.03-0.04 MPa, and after cultivation 48h, aseptic technique fills into three grade fermemtation tank; Three grade fermemtation tank temperature is by 28 ℃ of controls, and rotating speed is 160 r/min, and sterile air flow control is at 0.5-0.8 vvm, and tank pressure is controlled at 0.03-0.05 MPa, and culture cycle is 136 h;

2 supplying technicses

The fermentation culture cycle, by final concentration be the oxalate of 0.2 g/L, the soya-bean oil of 5 mL/L is aseptic filled in three grade fermemtation tank when 60 h.

Embodiment 2

1 fermentation culture

With embodiment 1

2 supplying technicses

The fermentation culture cycle, by final concentration be the oxalate of 0.5 g/L, the soya-bean oil of 2 mL/L is aseptic filled in three grade fermemtation tank when 80 h.

Embodiment 3

1 fermentation culture

With embodiment 1

2 supplying technicses

The fermentation culture cycle, by final concentration be the oxalate of 0.35 g/L, the soya-bean oil of 3 mL/L is aseptic filled in three grade fermemtation tank when 70 h.

Claims (1)

1. improve a method for Rifamycin Sodium fermentation yield, its operation steps is as follows:

(1) fermentation culture

Press seed and fermentative medium formula configuration nutrient solution, after moist heat sterilization, be cooled to 28 ℃, by the inoculum size of 3%-5%, bottle seed liquor of closing of Amycolatopsis mediterranei is accessed to a first class seed pot; First class seed pot temperature control is at 28 ℃, and sterile air flow is controlled by 0.8 vvm, and mixing speed is 160 r/min, and tank pressure is controlled at 0.03-0.04 MPa, and after cultivation 48h, aseptic technique fills into secondary seed tank; The temperature control of secondary seed tank is at 28 ℃, and sterile air flow is controlled by 0.8 vvm, and mixing speed is 160 r/min, and tank pressure is controlled at 0.03-0.04 MPa, and after cultivation 48h, aseptic technique fills into three grade fermemtation tank; Three grade fermemtation tank temperature is by 28 ℃ of controls, and rotating speed is 160 r/min, and sterile air flow control is at 0.5-0.8 vvm, and tank pressure is controlled at 0.03-0.05 MPa, and culture cycle is 136 h;

(2) supplying technics

Three grade fermemtation culture cycle is when 60-80 h, and by final concentration, being that the oxalate of 0.2-0.5 g/L and the soya-bean oil of 2-5 mL/L are aseptic fills in three grade fermemtation tank, can improve Rifamycin Sodium output.