Functionalized polyhydroxyalkanoate nano-beads as a stable biocatalyst for cost-effective production of the rare sugar d-allulose

Ran, Ganqiao; Tan, Dan; Zhao, Jiping; Fan, Fan; Zhang, Qiang; Wu, Xueyuan; Fan, Peiyao; Fang, Xinlei; Lu, Xiaoyun

doi:10.1016/j.biortech.2019.121673

Cited by 35 publications

(24 citation statements)

References 43 publications

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“…ClearColi technology produces a modified LPS, named Lipid IVA, that does not trigger the endotoxic response in human cells. Another ClearColi strain, BL21(DE3), has been increasingly used in the production of proteins, [25][26][27] as well as virus-like particles for vaccines. 28 To our knowledge, this is the first time that the ClearColi technology has been used to produce low-endotoxin rAAV vectors.…”

Section: Discussionmentioning

confidence: 99%

Low endotoxin E. coli strain-derived plasmids reduce rAAV vector-mediated immune responses both in vitro and in vivo

Zheng

Wang

Zhu³

et al. 2021

Molecular Therapy - Methods & Clinical Development

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The major challenge of recombinant adeno-associated virus (rAAV) vectors is host immunological barriers. Compared to the neutralizing antibody and the cytotoxic T lymphocyte response, the host immune responses induced by unsatisfactory rAAV manufacturing were largely ignored previously. rAAV vector production usually requires large amounts of plasmid DNAs. The DNA are commonly isolated from the DH5a bacterial strain, which contains lipopolysaccharide (LPS) contamination. LPS, also named endotoxin, in plasmid DNA is intractable, and residual endotoxin in the subsequent rAAV vectors may result in substantial host immune response. Recently, a ClearColi K12 bacterial strain is commercially available, with genetically modified LPS that does not trigger endotoxic response in mammalian cells. Here, we produced rAAV-DJ vectors by plasmids yielded from either DH5a or ClearColi K12 bacterial strains. Our data indicated that the ClearColi K12 strain had satisfactory protection for the rAAV inverted terminal repeat (ITR) sequence. As expected, the ClearColi K12derived rAAV-DJ vectors had lower endotoxin levels. The physical and biological equivalency of the purified viral stocks were confirmed by electron micrographs, Coomassie blue staining, and transduction assays. Most importantly, the ClearColi K12-derived rAAV-DJ vectors triggered reduced nuclear factor-kappa B (NF-kB) signaling pathway both in cell cultures in vitro and in C57BL/6 mice retinas in vivo. We believe that the use of the ClearColi K12 bacterial strain could eliminate the LPS in the purified vector stock at the source. Our data indicate its promising use in future clinical development.

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Section: Discussionmentioning

confidence: 99%

Low endotoxin E. coli strain-derived plasmids reduce rAAV vector-mediated immune responses both in vitro and in vivo

Zheng

Wang

Zhu³

et al. 2021

Molecular Therapy - Methods & Clinical Development

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“…Due to the cytotoxic nature of the cross-linking agent glutaraldehyde, safer cross-linking agents are needed for the production of immobilized d -allulose. Furthermore, functionalized polyhydroxyalkanoate nano-beads were used for the immobilization of enzymes, for which the products had a high bioactivity in complex reaction systems [ 134 ].…”

Section: Enzymatic Biotransformation Of D -Allulosementioning

confidence: 99%

Research Advances of d-allulose: An Overview of Physiological Functions, Enzymatic Biotransformation Technologies, and Production Processes

Xia

Cheng

et al. 2021

Foods

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d-allulose has a significant application value as a sugar substitute, not only as a food ingredient and dietary supplement, but also with various physiological functions, such as improving insulin resistance, anti-obesity, and regulating glucolipid metabolism. Over the decades, the physiological functions of d-allulose and the corresponding mechanisms have been studied deeply, and this product has been applied to various foods to enhance food quality and prolong shelf life. In recent years, biotransformation technologies for the production of d-allulose using enzymatic approaches have gained more attention. However, there are few comprehensive reviews on this topic. This review focuses on the recent research advances of d-allulose, including (1) the physiological functions of d-allulose; (2) the major enzyme families used for the biotransformation of d-allulose and their microbial origins; (3) phylogenetic and structural characterization of d-allulose 3-epimerases, and the directed evolution methods for the enzymes; (4) heterologous expression of d-allulose ketose 3-epimerases and biotransformation techniques for d-allulose; and (5) production processes for biotransformation of d-allulose based on the characterized enzymes. Furthermore, the future trends on biosynthesis and applications of d-allulose in food and health industries are discussed and evaluated in this review.

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“…Given the encouraging proof-of-concept results adapting the PHA particle technology for development of immobilized enzymes for uses in the food industry (Moldes et al, 2004;Rasiah and Rehm, 2009;Ran et al, 2017Ran et al, , 2019, production of commodity chemicals (Jahns and Rehm, 2015;Yang et al, 2015;Seo et al, 2016), production of fine chemicals Chen S. Y. et al, 2014;Tan et al, 2019), and bioremediation (Robins et al, 2013;Hooks and Rehm, 2015;Li et al, 2019), it is anticipated that research prototypes will be developed into industrial products.…”

Section: Potential Industrial Applications Of the Pha Particle Technomentioning

confidence: 99%

Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

Wong

Ogura

Chen

et al. 2020

Front. Bioeng. Biotechnol.

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Enzymes function as biocatalysts and are extensively exploited in industrial applications. Immobilization of enzymes using support materials has been shown to improve enzyme properties, including stability and functionality in extreme conditions and recyclability in biocatalytic processing. This review focuses on the recent advances utilizing the design space of in vivo self-assembled polyhydroxyalkanoate (PHA) particles as biocatalyst immobilization scaffolds. Self-assembly of biologically active enzyme-coated PHA particles is a one-step in vivo production process, which avoids the costly and laborious in vitro chemical cross-linking of purified enzymes to separately produced support materials. The homogeneous orientation of enzymes densely coating PHA particles enhances the accessibility of catalytic sites, improving enzyme function. The PHA particle technology has been developed into a remarkable scaffolding platform for the design of cost-effective designer biocatalysts amenable toward robust industrial bioprocessing. In this review, the PHA particle technology will be compared to other biological supramolecular assembly-based technologies suitable for in vivo enzyme immobilization. Recent progress in the fabrication of biological particulate scaffolds using enzymes of industrial interest will be summarized. Additionally, we outline innovative approaches to overcome limitations of in vivo assembled PHA particles to enable finetuned immobilization of multiple enzymes to enhance performance in multi-step cascade reactions, such as those used in continuous flow bioprocessing.

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Functionalized polyhydroxyalkanoate nano-beads as a stable biocatalyst for cost-effective production of the rare sugar d-allulose

Cited by 35 publications

References 43 publications

Low endotoxin E. coli strain-derived plasmids reduce rAAV vector-mediated immune responses both in vitro and in vivo

Low endotoxin E. coli strain-derived plasmids reduce rAAV vector-mediated immune responses both in vitro and in vivo

Research Advances of d-allulose: An Overview of Physiological Functions, Enzymatic Biotransformation Technologies, and Production Processes

Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

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