Rapid Fermentation Process for Dexotrolactic Acid

Abstract

The submerged cultivation of Rhizopus oryzae in rotary aluminum fermenters under stated conditions results in the fermentation of 13 per cent glucose solutions in 30 to 35 hours, with 70 to 75 per cent yields of dextrolactic acid. A carbon balance shows that, in addition to lactic acid and alcohol, an unidentified soluble material is produced equivalent to 7.4 per cent of the glucose consumed. Possible industrial advantages of this process are discussed.HE production of dextrolactic acid [also known as sarcolactic acid or I(+)lactic acid] by Rhizopus oryzm when cultivated on the surface of glucose nutrient solutions was the subject of previous communications from this division (6, 9). Surface cultivation gave 65 to 67 per cent yields of lactic acid in an incubation period of approximately 2 weeks. The present paper describes a process which involves the cultivation of this fungus in a submerged condition, whereby 13 per cent glucose solutions are fermented in 30 to 35 hours, with 70 to 75 per cent yields of d-lactic acid. This work is dependent on the use of the previously described rotary aluminum fermenters (4) which have also been employed as culture vessels to bring about the extremely rapid conversion of glucose to gluconic acid, using the organism Aspergillus niger (6, l l ) , and of sorbitol to sorbose, using the organism Acetobacter suboxydans (12). Materials and Methods TThe organism used in these studies was the previously described strain of Rhizopus oryzue Went and Geerligs (6), which was one of the best d-lactic said producers observed in the surface-growth studies. Although it is usually cultured on the surface of unagitated aedia, its growth and biochemical activity are enharxed when it is cultivated submerged under the favorable conditions described below.The culture of the organism in this work proceeded through the following three stages: (a) production ol' spores by growing on moist white bread for about 2 weeks, (b) germination of the spores in a glass bottle shaken during a period of 24 hours, and (c) the main fermentation, conducted in the rotary aluminum fermenters.The moist white bread cultures (one-half slice of bread in 200-cc. Erlenmeyer flasks) were inoculated from agar slants or from other bread cultures. After 5 to 6 days sporulation was profuse, and a t the age of about 2 weeks a portion of these spores was aseptically transferred to 50-75 cc. of sterile water and shaken to form a suspension, and a sample was removed to determine the spore concentration by means of a cytometer. A volume containing 420 million spores was then uised to inoculate 1.5 liters of germination medium. Usually this number of spores could be obtained from 5 to 10 cc. of spore suspension, and one bread culture would yield 5 to 10 billion spores.The germination medium had the following composition, in grams: Commercial glucose 110 KHzPO4 0.60 Urea 2 . 0 ZnSOa.7HzO 0.088 MgSO4.7HzO 0 . 2 5 CaCOa 10.0 Distilled water t o make 1000 oc.All of these components were sterilized together. The glucose was a commercial g...

Mass production of spores of lactic acid‐producing Rhizopus oryzae NBRC 5384 on agar plate

“…Seven‐ or eight‐day cultivation gave the maximum value and a longer cultivation than this decreased the spore numbers (data not shown). This outcome is consistent with the other reports . Spores were harvested on the 7th day in subsequent experiments unless otherwise stated.…”

“…The moisture (measured by a moisture meter) in the incubator was kept at 85-90% throughout the culture. The temperature (30 C) was the most frequently applied 2,4,6,9,10,13,14,18 although some other researchers applied other temperatures ranging from 25 to 35 C. 7,11,15,17 Other studies have no description of the incubation temperatures. 5,8,12,16…”

Organic Acid Production by Filamentous Fungi

“…was reported early in the last century (Ehrlich et al, 1911). However, the first report of an efficient submerged fermentation for the fungal production of L-lactic acid was in 1936 (Lockwood et al, 1936;Ward et al, 1938). This was the era in which the efficiencies of submerged fungal fermentations first became widely recognized.…”

“…This was the era in which the efficiencies of submerged fungal fermentations first became widely recognized. Ward et al (1938) described a fermentation process utilizing the Zygomycete genera, Rhizopus and Actinomucor in general, and R. oryzae (syn. arrhizus) specifically, which resulted in 63-69% yields of L-lactic acid from chemically defined media containing 15% glucose.…”

Studies of the Nutrition and Metabolism of the Aquatic Phycomycete, Allomyces

“…The mycelium was harvested by filtration, dried and weighed, and 50 per cent of the weight was assumed to be carbon according to the finding of Ward et al (1938): An aliquot of the culture filtrate was cleared of protein by the zinc sulfate method (Somogyi, 1945), acidified, and extracted with ether in a continuous liquid-liquid extractor for 24 hr. The mycelium was harvested by filtration, dried and weighed, and 50 per cent of the weight was assumed to be carbon according to the finding of Ward et al (1938): An aliquot of the culture filtrate was cleared of protein by the zinc sulfate method (Somogyi, 1945), acidified, and extracted with ether in a continuous liquid-liquid extractor for 24 hr.…”