Lysine production from methanol at 50°C using Bacillus methanolicus: Modeling volume control, lysine concentration, and productivity using a three‐phase continuous simulation

Lee, Grace H.; Hur, Won; Bremmon, Craig E.; Flickinger, Michael C.

doi:10.1002/(sici)1097-0290(19960320)49:6<639::aid-bit5>3.3.co;2-0

Cited by 6 publications

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“…HPS and PHI activities were assayed at 30°C as described previously[2]. The amount of protein was determined using a Bio‐Rad protein assay kit with bovine serum albumin as the standard.…”

Section: Methodsmentioning

confidence: 99%

“…The Bacillus strains belonging to the third group are distinct from the others in several respects, their growth ability at higher temperature, the methanol oxidation system catalyzed by NAD + ‐dependent methanol dehydrogenase[1], etc. Also, Bacillus strains are attractive for application to the production of useful compounds[2]. Genetic studies on the RuMP pathway have been performed with two methylotrophic bacteria belonging to the former two groups, Methylomonas aminofaciens 77a[3] and Mycobacterium gastri MB19[4], respectively.…”

Section: Introductionmentioning

confidence: 99%

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The ribulose monophosphate pathway operon encoding formaldehyde fixation in a thermotolerant methylotroph,Bacillus brevisS1

Yurimoto

Hirai

Yasueda

et al. 2002

FEMS Microbiology Letters

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The hps and phi genes encoding 3-hexulose-6-phosphate synthase and 6-phospho-3-hexuloisomerase, the key enzymes of the ribulose monophosphate (RuMP) pathway for formaldehyde fixation, were cloned from the chromosomal DNA of a thermotolerant methylotroph, Bacillus brevis S1. Enzyme induction and Northern blot analyses revealed that both the hps and phi genes are induced by methanol or ethanol, and that their expression is controlled polycistronically at the transcription stage. Sequence analysis also suggested that the hps and phi genes constitute an RuMP operon. The gene organization of the RuMP operon and its surrounding region are unique among bacteria possessing the RuMP pathway genes.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The ribulose monophosphate pathway operon encoding formaldehyde fixation in a thermotolerant methylotroph,Bacillus brevisS1

Yurimoto

Hirai

Yasueda

et al. 2002

FEMS Microbiology Letters

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Expression of recombinant green fluorescent protein in Bacillus methanolicus

Nilasari

Dover

Rech

et al. 2012

Biotechnology Progress

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Microbial biocatalysts are used in a wide range of industries to produce large scale quantities of proteins, amino acids, and commodity chemicals. While the majority of these processes use glucose or other low-cost sugars as the substrate, Bacillus methanolicus is one example of a biocatalyst that has shown sustained growth on methanol as a carbon source at elevated temperature (50-53°C optimum) resulting in reduced feed and utility costs. Specifically, the complete chemical process enabled by this approach takes methane from natural gas, and following a low-cost conversion to methanol, can be used for the production of high value products. In this study, production of recombinant green fluorescent protein (GFPuv) by B. methanolicus is explored. A plasmid was constructed that incorporates the methanol dehydrogenase (mdh) promoter of B. methanolicus MGA3 together with the GFPuv gene. The plasmid, pNW33N, was shown to be effective for expression in other Bacillus strains, although not previously in B. methanolicus. A published electroporation protocol for transformation of B. methanolicus was modified to result in expression of GFP using plasmid pNW33N-mdh-GFPuv (pNmG). Transformation was confirmed by both agarose gel electrophoresis and by observation of green fluorescence under UV light exposure. The mass yield of cells and protein were measured in shake flask experiments. The optimum concentration of methanol for protein production was found to be at 200 mM. Higher concentrations than 200 mM resulted in slightly higher biomass production but lower amounts of recombinant protein.

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Lysine Fermentation: History and Genome Breeding

Ikeda

2016

Amino Acid Fermentation

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Lysine fermentation by Corynebacterium glutamicum was developed in 1958 by Kyowa Hakko Kogyo Co. Ltd. (current Kyowa Hakko Bio Co. Ltd.) and is the second oldest amino acid fermentation process after glutamate fermentation. The fundamental mechanism of lysine production, discovered in the early stages of the process's history, gave birth to the concept known as "metabolic regulatory fermentation," which is now widely applied to metabolite production. After the development of rational metabolic engineering, research on lysine production first highlighted the need for engineering of the central metabolism from the viewpoints of precursor supply and NADPH regeneration. Furthermore, the existence of active export systems for amino acids was first demonstrated for lysine in C. glutamicum, and this discovery has resulted in the current recognition of such exporters as an important consideration in metabolite production. Lysine fermentation is also notable as the first process to which genomics was successfully applied to improve amino acid production. The first global "genome breeding" strategy was developed using a lysine producer as a model; this has since led to new lysine producers that are more efficient than classical industrial producers. These advances in strain development technology, combined with recent systems-level approaches, have almost achieved the optimization of entire cellular systems as cell factories for lysine production. In parallel, the continuous improvement of the process has resulted not only in fermentation processes with reduced load on downstream processing but also in commercialization of various product forms according to their intended uses. Nowadays lysine fermentation underpins a giant lysine demand of more than 2 million metric tons per year.

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Lysine production from methanol at 50°C using Bacillus methanolicus: Modeling volume control, lysine concentration, and productivity using a three‐phase continuous simulation

Cited by 6 publications

References 0 publications

The ribulose monophosphate pathway operon encoding formaldehyde fixation in a thermotolerant methylotroph,Bacillus brevisS1

The ribulose monophosphate pathway operon encoding formaldehyde fixation in a thermotolerant methylotroph,Bacillus brevisS1

Expression of recombinant green fluorescent protein in Bacillus methanolicus

Lysine Fermentation: History and Genome Breeding

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