Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris

Bumrungtham, Pornsiri; Promdonkoy, Peerada; Prabmark, Kanoknart; Bunterngsook, Benjarat; Boonyapakron, Katewadee; Tanapongpipat, Sutipa; Champreda, Verawat; Runguphan, Weerawat

doi:10.3390/jof8080767

Cited by 10 publications

(4 citation statements)

References 39 publications

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“…However, the method used in the present study reduces the number of steps required for isobutanol production, which would be expected to reduce production costs, making it a potentially effective approach to producing industrial isobutanol from plant biomass. Pichia pastoris PPY0311 Yes Sugarcane trash 3 steps 0.0482 ± 0.00170 0.000669 ± 0.0000236 [38] * The value was calculated from the concentration after 72 h of incubation.…”

Section: Comparison Of Isobutanol Productivitymentioning

confidence: 99%

Simultaneous Saccharification and Fermentation for Isobutanol Production from Banana Peel

Akita,

Shibata,

Komoriya

et al. 2024

Fermentation

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Each year, near 40 million tons of banana peels are discarded around the world. This plant biomass could potentially be utilized for energy production. Simultaneous saccharification and fermentation (SSF) is an effective method for producing biofuels from plant biomasses. Since SSF with enzymatic hydrolysis and fermentation are performed simultaneously in the same reactor, the production process is simpler than most existing methods. Here, we describe isobutanol production using SSF with hydrothermally treated banana peel samples and an Escherichia coli strain able to utilize glucose and xylose to produce isobutanol. To enhance the glucose and xylose concentrations, the reaction conditions for the enzymatic hydrolysis of plant biomass using two kinds of saccharification enzymes were optimized, including the enzyme unit ratio, reaction temperature and sample gram. When the optimized conditions for enzymatic hydrolysis were applied to SSF, the glucose and xylose produced from the hydrothermally treated samples were consumed, producing isobutanol. Moreover, the isobutanol concentration increased with an increasing initial culture pH, reaching 1.27 g/L at pH 6.5, which was consistent with the optimal initial culture pH for isobutanol production by this E. coli strain. Taken together, these results indicate that the established method is potentially useful for industrial isobutanol production.

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Section: Comparison Of Isobutanol Productivitymentioning

confidence: 99%

Simultaneous Saccharification and Fermentation for Isobutanol Production from Banana Peel

Akita,

Shibata,

Komoriya

et al. 2024

Fermentation

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“…The yeast strains were constructed from P. pastoris KM71 (Invitrogen), as shown in Table 1. The plasmids were derived from the pGAPHisKlARS and pGAPHyg vectors [18,29], as well as the commercially available pGAPZ vector (Invitrogen). The primers used are listed in Table S1.…”

Section: Media Strains and Transformationmentioning

confidence: 99%

Engineering Flocculation for Improved Tolerance and Production of d-Lactic Acid in Pichia pastoris

Sae-Tang

Bumrungtham

Mhuantong

et al. 2023

JoF

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d-lactic acid, a chiral organic acid, can enhance the thermal stability of polylactic acid plastics. Microorganisms such as the yeast Pichia pastoris, which lack the natural ability to produce or accumulate high amounts of d-lactic acid, have been metabolically engineered to produce it in high titers. However, tolerance to d-lactic acid remains a challenge. In this study, we demonstrate that cell flocculation improves tolerance to d-lactic acid and increases d-lactic acid production in Pichia pastoris. By incorporating a flocculation gene from Saccharomyces cerevisiae (ScFLO1) into P. pastoris KM71, we created a strain (KM71-ScFlo1) that demonstrated up to a 1.6-fold improvement in specific growth rate at high d-lactic acid concentrations. Furthermore, integrating a d-lactate dehydrogenase gene from Leuconostoc pseudomesenteroides (LpDLDH) into KM71-ScFlo1 resulted in an engineered strain (KM71-ScFlo1-LpDLDH) that could produce d-lactic acid at a titer of 5.12 ± 0.35 g/L in 48 h, a 2.6-fold improvement over the control strain lacking ScFLO1 expression. Transcriptomics analysis of this strain provided insights into the mechanism of increased tolerance to d-lactic acid, including the upregulations of genes involved in lactate transport and iron metabolism. Overall, our work represents an advancement in the efficient microbial production of d-lactic acid by manipulating yeast flocculation.

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“…Another P. pastoris metabolic engineering study targeted direct isobutanol production from sugarcane trash hydrolysate [ 119 ]. P. pastoris was engineered for heterologous xylose isomerase and endogenous xylulokinase overexpression to consume both C5 and C6 sugars in biomass.…”

Section: Introductionmentioning

confidence: 99%

“…For isobutanol production, endogenous amino acid biosynthetic pathway and 2-keto acid degradation pathway were overexpressed. Resulting strain produced isobutanol at a titer of 48.2 ± 1.7 mg.L −1 directly from minimal medium containing sugarcane trash hydrolysates as the sole carbon source [ 119 ].…”

Section: Introductionmentioning

confidence: 99%

Second generation Pichia pastoris strain and bioprocess designs

Ergün

Laçın

Çaloğlu

et al. 2022

Biotechnol Biofuels

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Yeast was the first microorganism used by mankind for biotransformation processes that laid the foundations of industrial biotechnology. In the last decade, Pichia pastoris has become the leading eukaryotic host organism for bioproduct generation. Most of the P. pastoris bioprocess operations has been relying on toxic methanol and glucose feed. In the actual bioeconomy era, for sustainable value-added bioproduct generation, non-conventional yeast P. pastoris bioprocess operations should be extended to low-cost and renewable substrates for large volume bio-based commodity productions. In this review, we evaluated the potential of P. pastoris for the establishment of circular bioeconomy due to its potential to generate industrially relevant bioproducts from renewable sources and waste streams in a cost-effective and environmentally friendly manner. Furthermore, we discussed challenges with the second generation P. pastoris platforms and propose novel insights for future perspectives. In this regard, potential of low cost substrate candidates, i.e., lignocellulosic biomass components, cereal by-products, sugar industry by-products molasses and sugarcane bagasse, high fructose syrup by-products, biodiesel industry by-product crude glycerol, kitchen waste and other agri-food industry by products were evaluated for P. pastoris cell growth promoting effects and recombinant protein production. Further metabolic pathway engineering of P. pastoris to construct renewable and low cost substrate utilization pathways was discussed. Although, second generation P. pastoris bioprocess operations for valorisation of wastes and by-products still in its infancy, rapidly emerging synthetic biology tools and metabolic engineering of P. pastoris will pave the way for more sustainable environment and bioeconomy. From environmental point of view, second generation bioprocess development is also important for waste recycling otherwise disposal of carbon-rich effluents creates environmental concerns. P. pastoris high tolerance to toxic contaminants found in lignocellulosic biomass hydrolysate and industrial waste effluent crude glycerol provides the yeast with advantages to extend its applications toward second generation P. pastoris strain design and bioprocess engineering, in the years to come. Graphical Abstract

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Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris

Cited by 10 publications

References 39 publications

Simultaneous Saccharification and Fermentation for Isobutanol Production from Banana Peel

Simultaneous Saccharification and Fermentation for Isobutanol Production from Banana Peel

Engineering Flocculation for Improved Tolerance and Production of d-Lactic Acid in Pichia pastoris

Second generation Pichia pastoris strain and bioprocess designs

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