Expression of unique chimeric human papilloma virus type 16 (HPV‐16) L1‐L2 proteins in <i>Pichia pastoris</i> and <i>Hansenula polymorpha</i>

Bredell, Helba; Smith, Jacques J.; Görgens, Johann F.; Zyl, Willem H. van

doi:10.1002/yea.3318

Cited by 29 publications

(12 citation statements)

References 82 publications

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“…The methylotrophic yeasts, e.g. K. phaffii , O. polymorpha and O. thermomethanolica, are typically used for heterologous protein production, due to their high-efficient heterogeneous protein secretion and glycosylation [8–10]. Kluyveromyces lactis is widely used in food and feed industries because of its ability to metabolize lactose and high protein secretion [11].…”

Section: Introductionmentioning

confidence: 99%

CRISPR-mediated genome editing in non-conventional yeasts for biotechnological applications

2019

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Non-conventional yeasts are playing important roles as cell factories for bioproduction of biofuels, food additives and proteins with outstanding natural characteristics. However, the precise genome editing is challenging in non-conventional yeasts due to lack of efficient genetic tools. In the past few years, CRISPR-based genome editing worked as a revolutionary tool for genetic engineering and showed great advantages in cellular metabolic engineering. Here, we review the current advances and barriers of CRISPR–Cas9 for genome editing in non-conventional yeasts and propose the possible solutions in enhancing its efficiency for precise genetic engineering.

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Section: Introductionmentioning

confidence: 99%

CRISPR-mediated genome editing in non-conventional yeasts for biotechnological applications

2019

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“…Various reports claim that higher recombinant protein production levels are obtained using MutS than when using Mut+ (Ascacio‐Martínez & Barrera‐Saldaña, ; Bredell, Smith, Görgens, & van Zyl, ; Cos et al, ; Krainer et al, ; Orman, Çalık, & Özdamar, ; Pla et al, ). However, the relatively few reports directly comparing Mut+ with MutS usually only focus on the final product yields and sometimes production rates and less on the reasons behind the observed differences.…”

Section: Introductionmentioning

confidence: 99%

Expression of recombinant enhanced green fluorescent protein provides insight into foreign gene‐expression differences between Mut+ and MutS strains of Pichia pastoris

Theron

Berríos

Steels

et al. 2019

Yeast

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Pichia pastoris is a very popular yeast for recombinant protein production, mainly due to the strong, methanol-inducible P AOX1 promoter. Methanol induction however poses several drawbacks. One approach to improve processes is to use MutS strains with reduced methanol catabolic ability. Various reports claim that MutS allows higher recombinant protein production levels than Mut+ but scarcely elaborate on reasons for differences. In this study, enhanced green fluorescent protein was used as a P AOX1 -driven reporter for the investigation of expression differences between Mut+ and MutS strains. Mut+ exhibited higher responses to methanol, with faster growth (0.07 vs. 0.01 hr −1 ) and higher consumption of methanol (2.25 vs.1.81 mmol/g DCW .hr) and oxygen (2.2 vs. 0.66 mmol/g DCW .hr) than MutS. Mut+ yielded more biomass than MutS (2.3 vs. 1.3 g DCW /L), and carbon dioxide analysis of bioreactor off-gas suggested that considerable amounts of methanol were consumed by Mut+ via the dissimilatory pathway. In contrast, it was demonstrated that the MutS population switched to an induced state more rapidly than Mut+. In addition, MutS exhibited 3.4-fold higher fluorescence levels per cell (77,509 vs. 23,783 SFU) indicative of higher recombinant protein production. The findings were verified by similar results obtained during the expression of a lipase. Based on the differences in response to methanol versus recombinant protein production, it was proposed that higher energy availability occurs in MutS for recombinant protein synthesis, contrary to Mut+ that uses the energy to maintain high levels of methanol catabolic pathways and biomass production.

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“…The inducible expression of OM-PLA1-engineered P. pastoris GS115 was investigated under different methanol concentrations and induction times (Bredell et al, 2018). Fermentation media were prepared by yeast extract 1% (w/v), peptone 2% (w/v), 100 mM pH6 potassium phosphate buffer, YNB 1.34% (w/v), biotin 4 × 10 -5 % (w/v), and glycerol 1% (w/v).…”

Section: Methodsmentioning

confidence: 99%

“…A Ni 2+ -chelating affinity chromatography column (Amersham Pharmacia Biotech, United Kingdom) was pre-equilibrated by a pH 8 buffer prepared by 20 mM imidazole, 50 mM NaH 2 PO 4 , and 300 mM NaCl. After the recombinant OM-PLA1 was purified by the column (Wang et al, 2009), OM-PLA1 profile analysis was performed by SDS-PAGE (Bio-Rad, United States) (Bredell et al, 2018).…”

Section: Methodsmentioning

confidence: 99%

Recombinant Expression of Serratia marcescens Outer Membrane Phospholipase A (A1) in Pichia pastoris and Immobilization With Graphene Oxide-Based Fe3O4 Nanoparticles for Rapeseed Oil Degumming

Yang

Jiang

et al. 2019

Front. Microbiol.

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Enzymatic degumming is an effective approach to produce nutritional, safe, and healthy refined oil. However, the high cost and low efficiency of phospholipase limit the application of enzymatic degumming. In this study, an 879 bp outer membrane phospholipase A (A1) (OM-PLA1) gene encoding 292 amino acid residues was isolated from the genome of Serratia marcescens . The recombinant OM-PLA1 profile of appropriately 33 KDa was expressed by the engineered Pichia pastoris GS115. The OM-PLA1 activity was 21.2 U/mL with the induction of 1 mM methanol for 72 h. The expression efficiencies of OM-PLA1 were 0.29 U/mL/h and 1.06 U/mL/OD600. A complex of magnetic graphene oxide (MGO) and OM-PLA1 (MGO-OM-PLA1) was prepared by immobilizing OM-PLA1 with graphene oxide-based Fe 3 O 4 nanoparticles by cross-linking with glutaraldehyde. The content of phosphorus decreased to 5.1 mg/kg rapeseed oil from 55.6 mg/kg rapeseed oil with 0.02% MGO-OM-PLA1 (w/w) at 50°C for 4 h. MGO-OM-PLA1 retained 51.7% of the initial activity after 13 times of continuous recycling for the enzymatic degumming of rapeseed oil. This study provided an effective approach for the enzymatic degumming of crude vegetable oil by developing a novel phospholipase and improving the degumming technology.

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Expression of unique chimeric human papilloma virus type 16 (HPV‐16) L1‐L2 proteins in Pichia pastoris and Hansenula polymorpha

Cited by 29 publications

References 82 publications

CRISPR-mediated genome editing in non-conventional yeasts for biotechnological applications

CRISPR-mediated genome editing in non-conventional yeasts for biotechnological applications

Expression of recombinant enhanced green fluorescent protein provides insight into foreign gene‐expression differences between Mut+ and MutS strains of Pichia pastoris

Recombinant Expression of Serratia marcescens Outer Membrane Phospholipase A (A1) in Pichia pastoris and Immobilization With Graphene Oxide-Based Fe3O4 Nanoparticles for Rapeseed Oil Degumming

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