Increased Productivity and Antifreeze Activity of Ice-binding Protein from Flavobacterium frigoris PS1 Produced using Escherichia coli as Bioreactor

Kim, Eun Jae; Kim, Jung Eun; Hwang, Jisub; Kim, Il-Chan; Lee, Sung Gu; Kim, Sanghee; Lee, Jun Hyuck; Han, Se Jong

doi:10.1134/s0003683819050077

Cited by 7 publications

(5 citation statements)

References 31 publications

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“…Qualitative confirmation of antifreeze activity was based on changes in ice crystal morphology compared to those crystals grown in a 30% (w/v) solution of sucrose [34,36]. Measurement of TH activity was carried out according to the method of Patel and Graether [37] with some modifications, using an Otago nanoliter osmometer (Otago Osmometers, Dunedin, New Zealand). A 20 nL drop of apoplastic extract was inserted into a Cargille oil Type B droplet in the cold-stage sample holder.…”

Section: Antifreeze Activity Assaysmentioning

confidence: 99%

Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study

Short,

Zamorano,

Aranzaez-Ríos

et al. 2024

Biomolecules

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Antifreeze proteins (AFPs) are natural biomolecules found in cold-adapted organisms that lower the freezing point of water, allowing survival in icy conditions. These proteins have the potential to improve cryopreservation techniques by enhancing the quality of genetic material postthaw. Deschampsia antarctica, a freezing-tolerant plant, possesses AFPs and is a promising candidate for cryopreservation applications. In this study, we investigated the cryoprotective properties of AFPs from D. antarctica extracts on Atlantic salmon spermatozoa. Apoplastic extracts were used to determine ice recrystallization inhibition (IRI), thermal hysteresis (TH) activities and ice crystal morphology. Spermatozoa were cryopreserved using a standard cryoprotectant medium (C+) and three alternative media supplemented with apoplastic extracts. Flow cytometry was employed to measure plasma membrane integrity (PMI) and mitochondrial membrane potential (MMP) postthaw. Results showed that a low concentration of AFPs (0.05 mg/mL) provided significant IRI activity. Apoplastic extracts from D. antarctica demonstrated a cryoprotective effect on salmon spermatozoa, with PMI comparable to the standard medium. Moreover, samples treated with apoplastic extracts exhibited a higher percentage of cells with high MMP. These findings represent the first and preliminary report that suggests that AFPs derived from apoplastic extracts of D. antarctica have the potential to serve as cryoprotectants and could allow the development of novel freezing media.

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Section: Antifreeze Activity Assaysmentioning

confidence: 99%

Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study

Short,

Zamorano,

Aranzaez-Ríos

et al. 2024

Biomolecules

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“…In recent years, Kim et al . (2019) used E . coli as the host and introduced the pET28a‐FfIBP vector system, the recombinant E .…”

Section: Feasibility Of Large‐scale Production Of Afpsmentioning

confidence: 99%

“…Later, Tab et al (2017) also produced 39.5 mg L À1 AFP1 of Glaciozyma antarctica in a 5 L fermentation tank using Pichia pastoris. In recent years, Kim et al (2019) used E. coli as the host and introduced the pET28a-FfIBP vector system, the recombinant E. coli eventually produced 1.6 g L À1 FfIBP, the yield was greatly improved.…”

Section: Feasibility Of Large-scale Production Of Afpsmentioning

confidence: 99%

Recent advances, challenges and functional applications of antifreeze protein in food industry

Xiao,

Hu,

Zhu

et al. 2023

Int J of Food Sci Tech

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SummaryFreezing is a common food preservation method that can extend the shelf life of food, but the ice crystals and recrystallisation during freezing, storage and transportation cause significant damage to frozen food. Antifreeze proteins are a class of proteins that widely exist in organisms living in cold conditions, such as fish, insects, plants and microorganisms in Antarctica and other cold regions. Antifreeze proteins can reduce the freezing point of water, inhibit recrystallisation effects, modify ice crystal morphology and effectively suppress the quality deterioration caused by ice crystals during food freezing. This review comprehensively introduces the source, antifreeze mechanism, application in food, influencing factors and regulating methods of antifreeze activity of antifreeze proteins, as well as the limitations of antifreeze proteins. It also looks forward to the future application of antifreeze proteins in food.

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“…were previously found to harbor multiple copies of these genes (Liu et al, 2019). Ice-binding (IBP) or antifreeze (ATF) proteins inhibiting formation of ice crystals and their recrystallization were not found in either genome, although gene ffIBP encoding IBP was already detected in the genome of psychrophilic marine strain, Flavobacterium frigoris PS1 (Kim et al, 2019). As icenucleating activity was detected in flavobacteria strains found confined in ice cores (Christner et al, 2000;Wilson et al, 2006), it is likely that genes encoding IPBs and ATFs are present, however, these remain unassigned with hypothetical functions.…”

Section: Genomic Features Associated With Cold-adaptationmentioning

confidence: 99%

Flavobacterium flabelliforme sp. nov. and Flavobacterium geliluteum sp. nov., Two Multidrug-Resistant Psychrotrophic Species Isolated From Antarctica

et al. 2021

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Despite unfavorable Antarctic conditions, such as cold temperatures, freeze-thaw cycles, high ultraviolet radiation, dryness and lack of nutrients, microorganisms were able to adapt and surprisingly thrive in this environment. In this study, eight cold-adapted Flavobacterium strains isolated from a remote Antarctic island, James Ross Island, were studied using a polyphasic taxonomic approach to determine their taxonomic position. Phylogenetic analyses based on the 16S rRNA gene and 92 core genes clearly showed that these strains formed two distinct phylogenetic clusters comprising three and five strains, with average nucleotide identities significantly below 90% between both proposed species as well as between their closest phylogenetic relatives. Phenotyping revealed a unique pattern of biochemical and physiological characteristics enabling differentiation from the closest phylogenetically related Flavobacterium spp. Chemotaxonomic analyses showed that type strains P4023T and P7388T were characterized by the major polyamine sym-homospermidine and a quinone system containing predominantly menaquinone MK-6. In the polar lipid profile phosphatidylethanolamine, an ornithine lipid and two unidentified lipids lacking a functional group were detected as major lipids. These characteristics along with fatty acid profiles confirmed that these species belong to the genus Flavobacterium. Thorough genomic analysis revealed the presence of numerous cold-inducible or cold-adaptation associated genes, such as cold-shock proteins, proteorhodopsin, carotenoid biosynthetic genes or oxidative-stress response genes. Genomes of type strains surprisingly harbored multiple prophages, with many of them predicted to be active. Genome-mining identified biosynthetic gene clusters in type strain genomes with a majority not matching any known clusters which supports further exploratory research possibilities involving these psychrotrophic bacteria. Antibiotic susceptibility testing revealed a pattern of multidrug-resistant phenotypes that were correlated with in silico antibiotic resistance prediction. Interestingly, while typical resistance finder tools failed to detect genes responsible for antibiotic resistance, genomic prediction confirmed a multidrug-resistant profile and suggested even broader resistance than tested. Results of this study confirmed and thoroughly characterized two novel psychrotrophic Flavobacterium species, for which the names Flavobacterium flabelliforme sp. nov. and Flavobacterium geliluteum sp. nov. are proposed.

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Increased Productivity and Antifreeze Activity of Ice-binding Protein from Flavobacterium frigoris PS1 Produced using Escherichia coli as Bioreactor

Cited by 7 publications

References 31 publications

Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study

Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study

Recent advances, challenges and functional applications of antifreeze protein in food industry

Flavobacterium flabelliforme sp. nov. and Flavobacterium geliluteum sp. nov., Two Multidrug-Resistant Psychrotrophic Species Isolated From Antarctica

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