Improved oxygen transfer and increased l-lactic acid production by morphology control of Rhizopus oryzae in a static bed bioreactor

Rhizopus oryzae was immobilized on a cotton matrix in a static bed bioreactor. Compared with free cells in a stirred tank bioreactor, immobilized R. oryzae in this bioreactor gave higher lactic acid production but lower ethanol production. The highest lactic acid production rate (2.09 g/L h) with the final concentration of 37.83 g/L from 70 g/L glucose was achieved when operating the bioreactor at 700 rpm and 0.5 vvm air. To better understand the relationship between shear effects (agitation and aeration) and R. oryzae morphology and metabolism, oxygen transfer rate, fermentation kinetics, and lactate dehydrogenase activity were determined. In immobilized cell culture, higher oxygen transfer rate and lactic acid production were achieved but lower lactate dehydrogenase activity was found as compared with those in free cell culture operated at the same conditions. These results clearly imply that mass transport was the rate controlling step in lactic acid fermentation by R. oryzae.     

D-Lactic acid fermentation performance and the enzyme activity of a novel bacterium Terrilactibacillus laevilacticus SK5–6

Purpose

The aim of this study was to prove that Terrilactibacillus laevilacticus SK5-6, a novel D-lactate producer, exhibited a good fermentation performance comparing to the reference D-lactate producer Sporolactobacillus sp.

Methods

Glucose bioconversion for D-lactate production and the activity of five key enzymes including phosphofructokinase (PFK), pyruvate kinase (PYK), D-lactate dehydrogenase (D-LDH), L-lactate dehydrogenase (L-LDH), and lactate isomerase (LI) were investigated in the cultivation of T. laevilacticus SK5–6 and S. laevolacticus 0361^T.

Results

T. laevilacticus SK5–6 produced D-lactate at higher yield, productivity, and optical purity compared with S. laevolacticus 0361^T. T. laevilacticus SK5–6, the catalase-positive isolate, simultaneously grew and produced D-lactate without lag phase while delayed growth and D-lactate production were observed in the culture of S. laevolacticus 0361^T. The higher production of D-lactate in T. laevilacticus SK5–6 was due to the higher growth rate and the higher specific activities of the key enzymes observed at the early stage of the fermentation. The low isomerization activity was responsible for the high optical purity of D-lactate in the cultivation of T. laevilacticus SK5–6.

Conclusion

The lowest specific activity of PFK following by PYK and D/L-LDHs, respectively, indicated that the conversion of fructose-6-phosphate was the rate limiting step. Under the well-optimized conditions, the activation of D/L-LDHs by fructose-1,6-phosphate and ATP regeneration by PYK drove glucose bioconversion toward D-lactate. The optical purity of D-lactate was controlled by D/L-LDHs and the activation of isomerases. High D-LDH with limited isomerase activity was preferable during the fermentation as it assured the high optical purity.

     

Characterization of<Emphasis Type="SmallCaps"> D</Emphasis>-lactic acid,spore-forming bacteria and <Emphasis Type="Italic">Terrilactibacillus laevilacticus</Emphasis> SK5-6 as potential industrial strains

In this study, we screened and isolated D-lactic acid-producing bacteria from soil and tree barks collected in Thailand. Among the isolates obtained, Terrilactibacillus laevilacticus SK5-6 exhibited good D-lactate production in the primary screening fermentation (99.27 g/L final lactate titer with 0.90 g/g yield, 1.38 g/L?h, and 99.00% D-enantiomer equivalent). Terrilactibacillus laevilacticus SK5-6 is a Gram-positive, endospore-forming, homofermentative D-lactate producer that can ferment a wide range of sugars to produce D-lactate. Unlike the typical D-lactate producers, such as catalase-negative Sporolactobacillus sp., T. laevilacticus SK5-6 possesses catalase activity; therefore, a two-phase fermentation was employed for D-lactate production. During an aerobic preculture stage, a high-density cell mass was rapidly obtained due to aerobic respiration. When transferred to the fermentation stage at the correct physiological stage (inoculum age) and proper concentration of cell mass (inoculum size), T. laevilacticus rapidly converted glucose into D-lactate under anaerobic conditions, resulting in a high final lactate titer (102.22 g/L), high yield (0.84 g/g), and high productivity (2.13 g/L?h). When the process conditions were shifted from an aerobic to an anaerobic environment, unlike other lactate-producing bacteria, the mixed acid fermentation route was not activated in the culture of T. laevilacticus SK5-6 during the fermentation stage when some trace oxygen still remained. Our study demonstrates the excellent characteristics of this isolate for D-lactate production; in particular, a high product yield was obtained without byproduct formation. Based on these key characteristics of T. laevilacticus SK5-6, we suggest that this isolate is a novel D-lactate producer for use in industrial fermentation.     

A homofermentative Bacillus sp. BC-001 and its performance as a potential l-lactate industrial strain

Bioprocess and Biosystems Engineering - Bacillus sp. BC-001 was first reported as a potent thermotolerant and homofermentative strain for an industrial-scale l-lactate production. In a flask...     

A modified approach for high-quality RNA extraction of spore-forming Bacillus subtilis at varied physiological stages

Molecular Biology Reports - High quality RNA is required for the molecular study. Sample preparation of the spore-forming, Gram-positive bacteria like Bacillus sp., remains challenging although...