Influence of codon optimization, promoter, and strain selection on the heterologous production of a β-fructofuranosidase from Aspergillus fijiensis ATCC 20611 in Pichia pastoris

Coetzee, Gerhardt; Smith, Jacques J.; Görgens, Johann F.

doi:10.1007/s12223-022-00947-8

Cited by 5 publications

(17 citation statements)

References 53 publications

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“…It has been found in soil on the Fiji Islands and on Lactuca sative in Indonesia and in indoor air, but it has never been reported in China in the past. In addition, it has a substantial economic value, as it includes fermenters of foodstuffs and key cell factories for the production of β-fructofuranosidase [60,61].…”

Section: Discussionmentioning

confidence: 99%

First Record of Aspergillus fijiensis as an Entomopathogenic Fungus against Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae)

Yan

Líu

Idrees

et al. 2022

JoF

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The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae) is the most widespread and devastating pest species in citrus orchards and is the natural vector of the phloem-limited bacterium that causes Huanglongbing (HLB) disease. Thus, reducing the population of D. citri is an important means to prevent the spread of HLB disease. Due to the long-term use of chemical control, biological control has become the most promising strategy. In this study, a novel highly pathogenic fungal strain was isolated from naturally infected cadavers of adult D. citri. The species was identified as Aspergillus fijiensis using morphological identification and phylogenetic analysis and assigned the strain name GDIZM-1. Tests to detect aflatoxin B1 demonstrated that A. fijiensis GDIZM-1 is a non-aflatoxin B1 producer. The pathogenicity of the strain against D. citri was determined under laboratory and greenhouse conditions. The results of the laboratory study indicated that nymphs from the 1st to 5th instar and adults of D. citri were infected by A. fijiensis GDIZM-1. The mortality of nymphs and adults of D. citri caused by infection with A. fijiensis increased with the concentration of the conidial suspension and exposure time, and the median lethal concentration (LC50) and median lethal time (LT50) values gradually decreased. The mortality of D. citri for all instars was higher than 70%, with high pathogenicity at the 7th day post treatment with 1×108 conidia/mL. The results of the greenhouse pathogenicity tests showed that the survival of D. citri adults was 3.33% on the 14th day post-treatment with 1 × 108 conidia/mL, which was significantly lower than that after treatment with the Metarhizium anisopliae GDIZMMa-3 strain and sterile water. The results of the present study revealed that the isolate of A. fijiensis GDIZM-1 was effective against D. citri and it provides a basis for the development of a new microbial pesticide against D. citri after validation of these results in the field.

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Section: Discussionmentioning

confidence: 99%

First Record of Aspergillus fijiensis as an Entomopathogenic Fungus against Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae)

Yan

Líu

Idrees

et al. 2022

JoF

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“…The production of scFOS via the transfructosylation of sucrose present in sugarcane juice, achieved with an enzyme produced by a recombinant strain of Pichia pastoris yeast, was simulated. This simulation was based on experimental data reported by Coetzee et al 13 and Trollope et al 18 The performances of scFOS production methods when using concentrated sugarcane juice from a mechanical crusher were assumed to be similar to those proposed for intermediate sugarcane streams such as A-molasses. 1,13 The simulation for Scenario 1-s consisted of three sections: sugarcane pretreatment (crushing, juice clarification, and concentration), enzyme production, and scFOS production and purification.…”

Section: Scenario 1-s: Small-scale Production Of Short-chain Fructool...mentioning

confidence: 99%

“…This simulation was based on experimental data reported by Coetzee et al 13 and Trollope et al 18 The performances of scFOS production methods when using concentrated sugarcane juice from a mechanical crusher were assumed to be similar to those proposed for intermediate sugarcane streams such as A-molasses. 1,13 The simulation for Scenario 1-s consisted of three sections: sugarcane pretreatment (crushing, juice clarification, and concentration), enzyme production, and scFOS production and purification. During pretreatment, sugarcane was crushed in a crusher mill, simulated by a component separator.…”

Section: Scenario 1-s: Small-scale Production Of Short-chain Fructool...mentioning

confidence: 99%

“…19 The enzyme production stage included all the processes involved in the preparation of the enzyme inoculum, enzyme fermentation, and partial enzyme purification. 1,13 For this stage, a 1000 L bioreactor was simulated as an RYield unit in Aspen Plus by specifying the desired cell and protein yields, as previously reported. 13 The bioreactor was prepared with basal salt medium and sterilized with high-pressure steam.…”

Section: Scenario 1-s: Small-scale Production Of Short-chain Fructool...mentioning

confidence: 99%

“…10 However, ethanol production directly from sugarcane without juice extraction through the use of solid-state fermentation (SSF) has been demonstrated in an industrial pilot, 11 and is a more appropriate production method for small-scale biorefineries, based on reduced processing requirements. The availability of low-cost and energy-efficient processing methods, together with attractive market prices, 7,12 and suitability of production using lower purity substrate 13 Original Article: Processing of sugarcane to fructooligosaccharides and ethanol 1568 © 2023 The Authors. Biofuels, Bioproducts and Biorefining published by Society of Industrial Chemistry and John Wiley & Sons Ltd.…”

Section: Introductionmentioning

confidence: 99%

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Economic comparison of decentralized versus centralized processing of sugarcane to fructooligosaccharides and ethanol

Klaver

Petersen

Görgens

2023

Biofuels Bioprod Bioref

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The global sugarcane industry is under threat due to low international prices and oversupply. This requires new revenue opportunities through a diversification strategy. The economic competitiveness of small‐scale, decentralized biorefineries to produce short‐chain fructooligosaccharides (scFOS) and/or ethanol, both valuable products with growing market applications, in comparison to large‐scale, centralized facilities annexed to existing sugarcane mills, was therefore considered. The economic feasibility of three potential scenarios was investigated using AspenPlus process simulations of small‐ and large‐scale facilities. In Scenario 1 only scFOS was produced. In Scenario 2 only ethanol was produced, and in Scenario 3 scFOS and ethanol were co‐produced. The small‐scale implementation of Scenarios 1 and 3 was economically feasible, with minimum selling prices (MSPs) below market prices. However, in all scenarios the large‐scale implementation of technologies in facilities annexed to an existing sugar mill improved economic viability, despite efforts to minimize the disadvantages of small‐scale facilities and their capital costs through the selection of simplified process technologies and decentralized processing. Despite this, the small‐scale production of scFOS is the preferred investment option for scFOS production due to the very high scFOS production rates of the large‐scale scenarios, which could flood the market. Processing of freshly harvested sugarcane in decentralized, small‐scale facilities increased the feedstock cost per unit of sugar, in comparison with the use of molasses obtained by annexation to an existing sugar mill. Simplified sugarcane processing to ethanol also lowered the product yield and increased capital costs. Further improvements in process technology design and performance for small‐scale facilities are required to make these financially competitive with large‐scale facilities.

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Evaluation of different glycerol fed-batch strategies in a lab-scale bioreactor for the improved production of a novel engineered β-fructofuranosidase enzyme in Pichia pastoris

Coetzee,

García-Aparicio,

Bosman

et al. 2024

World J Microbiol Biotechnol

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The β-fructofuranosidase enzyme from Aspergillus niger has been extensively used to commercially produce fructooligosaccharides from sucrose. In this study, the native and an engineered version of the β-fructofuranosidase enzyme were expressed in Pichia pastoris under control of the glyceraldehyde-3-phosphate dehydrogenase promoter, and production was evaluated in bioreactors using either dissolved oxygen (DO-stat) or constant feed fed-batch feeding strategies. The DO-stat cultivations produced lower biomass concentrations but this resulted in higher volumetric activity for both strains. The native enzyme produced the highest volumetric enzyme activity for both feeding strategies (20.8% and 13.5% higher than that achieved by the engineered enzyme, for DO-stat and constant feed, respectively). However, the constant feed cultivations produced higher biomass concentrations and higher volumetric productivity for both the native as well as engineered enzymes due to shorter process time requirements (59 h for constant feed and 155 h for DO-stat feed). Despite the DO-stat feeding strategy achieving a higher maximum enzyme activity, the constant feed strategy would be preferred for production of the β-fructofuranosidase enzyme using glycerol due to the many industrial advantages related to its enhanced volumetric enzyme productivity.

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Influence of codon optimization, promoter, and strain selection on the heterologous production of a β-fructofuranosidase from Aspergillus fijiensis ATCC 20611 in Pichia pastoris

Cited by 5 publications

References 53 publications

First Record of Aspergillus fijiensis as an Entomopathogenic Fungus against Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae)

First Record of Aspergillus fijiensis as an Entomopathogenic Fungus against Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae)

Economic comparison of decentralized versus centralized processing of sugarcane to fructooligosaccharides and ethanol

Evaluation of different glycerol fed-batch strategies in a lab-scale bioreactor for the improved production of a novel engineered β-fructofuranosidase enzyme in Pichia pastoris

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