Discovery of novel highly active and stable aspartate dehydrogenases

Li, Hao; Zhu, Tong; Miao, Liangtian; Zhang, Dan; Li, Yongxian; Li, Qi; Li, Yin

doi:10.1038/s41598-017-05522-7

Cited by 9 publications

(7 citation statements)

References 18 publications

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“…P. pastoris possesses a mitochondrial AAT (encoded by AAT1) and a cytosolic AAT (AAT2), and both genes were cloned into the AOX1 vector and tested for their effects on β-alanine synthesis, respectively. In a previous paper, we reported that an AspDH from S. proteamaculans has high activity and stability (Li et al 2017a). Thus, the SpeAspDH was also overexpressed in recombinant P. pastoris using the same strategy as AAT1 and AAT2.…”

Section: Resultsmentioning

confidence: 97%

Metabolic Engineering of the Methylotrophic Yeast Pichia Pastoris (Komagataella phaffii) for the Production of Β-alanine From Methanol

Miao

Zhu

2021

Preprint

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β-Alanine (3-aminopropionic acid), is the only naturally occurring β-amino acid and an important precursor for the synthesis of a variety of nitrogen-containing chemicals. Fermentative production of β-alanine from renewable feedstocks such as glucose has attracted significant interest in recent years. Methanol has become an emerging and promising renewable feedstock for biomanufacturing as an alternative to glucose. In this work, we demonstrated the feasibility of β-alanine production from methanol using Pichia pastoris (Komagataella phaffii) as a methylotrophic cell factory. Aspartate decarboxylases (ADCs) from different sources were screened and expressed in P. pastoris, followed by the optimization of aspartate decarboxylation by increasing the ADC copy number and C4 precursor supply via the overexpression of aspartate dehydrogenase. The production potential of the best strain was further evaluated in a 1-liter fermenter, and a β-alanine titer of 5.6 g/L was obtained. To our best knowledge, this is the highest chemical production titer ever reached in P. pastoris using methanol as the substrate.

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Section: Resultsmentioning

confidence: 97%

Metabolic Engineering of the Methylotrophic Yeast Pichia Pastoris (Komagataella phaffii) for the Production of Β-alanine From Methanol

Miao

Zhu

2021

Preprint

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“…P. pastoris possesses a mitochondrial AAT (encoded by AAT1 ) and a cytosolic AAT ( AAT2 ), and both genes were cloned into the pMPICKmHis vector and tested for their effects on β-alanine synthesis, respectively. In a previous paper, we reported that an AspDH from S. proteamaculans has high activity and stability (Li et al 2017a ). Thus, the SpeAspDH was also overexpressed in recombinant P. pastoris using the same strategy as AAT1 and AAT2.…”

Section: Resultsmentioning

confidence: 97%

“…First, we aimed at increasing the conversion of OAA into aspartate by screening for appropriate enzymes for reductive amination. Although the overexpression of AAT was widely used to increase aspartate flux for the production of aspartate family amino acids (AFAAs) in industrial microbes such as C. glutamicum and E. coli (Li et al 2017a , b ; Piao et al 2019 ), the efficacy of AspDH overexpression for enhancing AFAAs production was rarely. The present study demonstrates that the overexpression of SpeAspDH can significantly increase β-alanine production, corroborating the potential of AspDHs for the production of AFAAs.…”

Section: Discussionmentioning

confidence: 99%

Metabolic engineering of methylotrophic Pichia pastoris for the production of β-alanine

Miao

Zhu

2021

Bioresour. Bioprocess.

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Abstractβ-Alanine (3-aminopropionic acid) is the only naturally occurring β-amino acid and an important precursor for the synthesis of a variety of nitrogen-containing chemicals. Fermentative production of β-alanine from renewable feedstocks such as glucose has attracted significant interest in recent years. Methanol has become an emerging and promising renewable feedstock for biomanufacturing as an alternative to glucose. In this work, we demonstrated the feasibility of β-alanine production from methanol using Pichia pastoris (Komagataella phaffii) as a methylotrophic cell factory. L-Aspartate-α-decarboxylases (ADCs) from different sources were screened and expressed in P. pastoris, followed by the optimization of aspartate decarboxylation by increasing the ADC copy number and C4 precursor supply via the overexpression of aspartate dehydrogenase. The production potential of the best strain was further evaluated in a 1-L fermenter, and a β-alanine titer of 5.6 g/L was obtained. To our best knowledge, this is the highest metabolite production titer ever reached in P. pastoris using methanol as the substrate. Graphic abstract

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“…Many organic matter sources can be used as fertilizer, namely manure, sewage sludge, plant stems, compost, biogas residue, biogas slurry [7]. The application of organic fertilizers is usually combined with inorganic fertilizers.…”

Section: Introductionmentioning

confidence: 99%

Application of Goat Manure Briquettes on Red Chili Cultivation in Coastal Sandy Soil

Budiyanto

Mulyono

Achmad

2022

IOP Conf. Ser.: Earth Environ. Sci.

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Application of organic matter could prevent a high infiltration problem commonly found in coastal sandy soil. Due to its slow-release properties, organic matter added to this sandy soil may improve the groundwater retention capacity. This study was aimed to investigate the capacity of goat manure briquettes (GMB) in comparison with goat manure to support the productivity of red chili cultivated in coastal sandy soil. The study was carried out in a completely randomized design consisting of 3 doses of goat manure briquettes (GMB) and goat manure (GM) applied (30, 35, 40 tons/ha) with 12 replications. Each dose was applied to the fertilized soils (400 kg/ha urea + 300 kg/ha SP-36 + 300 kg/ha KCl) then incubated for a week. These treated soils were then used for red chili cultivation. Plant growth and yield were monitored and data collected were statistically analysed using DNMRT with significance level of 5%. GMB application showed insignificant effect towards the chili performance in sandy soil. In contrast, GM application revealed higher plant performance and yield. Application of 40 tons/ha GM exhibited the highest fruit weight compared to other treatments. According to these results, the formulation of GM into briquette might reduce its capacity in supporting plant growth.

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Discovery of novel highly active and stable aspartate dehydrogenases

Cited by 9 publications

References 18 publications

Metabolic Engineering of the Methylotrophic Yeast Pichia Pastoris (Komagataella phaffii) for the Production of Β-alanine From Methanol

Metabolic Engineering of the Methylotrophic Yeast Pichia Pastoris (Komagataella phaffii) for the Production of Β-alanine From Methanol

Metabolic engineering of methylotrophic Pichia pastoris for the production of β-alanine

Application of Goat Manure Briquettes on Red Chili Cultivation in Coastal Sandy Soil

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