Pilot-Scale Batch Production of High Lysine Content Animal Feed Suppliment

Pilot-Scale Batch Production Of High Lysine Content Animal Feed Suppliment

This is a Fed-batch fermentation flow schematic for the production of high lysine content animal feed supplement from methanol using the methylotroph Bacillus methanolicus strain MGA3. The inoculum is started from a 1 ml vial of frozen vegetative rods. Seed build-up was accomplished by using 4-liter shake flasks overnight with minimal salts medium + methanol to give a 5% v/v starter culture for the next stage. The next stage is a New Brunswick Scientific (NBS) fermenter run overnight with dissolved O2 control and automatic methanol addition as needed, to provide a 10% v/v starter culture for the next stage. A 300-liter fermenter is used for the final fermentation production stage. It has automatic methanol addition, pH control by automatic addition of ammonium hydroxide, and dissolved oxygen control by proportional addition of oxygen to the sparge air by a mass flow controller. This ran for 20 h and yielded a 300 liter total volume, high lysine, high cell density product delivered for spray dryer processing (98, 112, 136, 190, 203, 216, 228, 230).


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Pilot-Scale Continuous Production of High Lysine Content Animal Feed Suppliment

Pilot-Scale Continuous Production Of High Lysine Content Animal Feed Suppliment

This is a continuous fermentation flow schematic for the production of high lysine content animal feed supplement from methanol using the methylotroph Bacillus methanolicus strain MGA3. The inoculum was started from a 1 ml vial of frozen vegetative rods. Seed build-up was accomplished by using 4-liter shake flasks overnight with minimal salts medium + methanol to give a 5% v/v starter culture for the next stage. The next stage was a New Brunswick Scientific (NBS) fermenter run overnight with dissolved O2 control and automatic methanol addition as needed, to provide a 10% v/v starter culture for the next stage.

The fermentation production stage consisted of a 150-liter Chemap reactor with a 100-liter working volume. At an optical density (OD620) of 75, continuous feeding was begun. The dilution rate was 2.3. The reactor was run at 800 RPM, with a 50 LPM air flow rate, proportional addition of oxygen to sparge air was used to control dissolved O2. Methanol was measured and controlled a by probe, controller, and pump combination. A constant volume of 100 liters was maintained by reactor load cells and an automatically controlled harvest valve. The automatic addition of anhydrous ammonia gas to the sparge air/oxygen line proved to be an effective method of pH control.

A Questor Process Mass Spectrometer (ABB Extrel Corp.) was used to measure inlet and exhaust gasses for oxygen, nitrogen, argon, methanol, ammonia, and water vapor. Oxygen uptake rates (OUR), carbon dioxide evolution rates (CER), and respiratory quotients (RQ) were continuously determined by a Fischer-Rosemount (RS3) process control system. Methanol and ammonia were continuously monitored by load cell platforms which relayed signals for methanol and anhydrous ammonia fed. This information was then used by the RS3 computers to determine carbon and nitrogen balances (68, 136, 190, 203, 216, 228, 230).


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Craig Bremmon
CEBTech Services
503 11 th Ave. SE, Suite 5
Aberdeen, SD 57401

bremm001@nvc.net
Phone: 605 226-3595
Mobile: 605 377-4062
Fax: 605 226-3595