Abstract
Thraustochytrids are heterotrophic fungus-like protists that can dissolve organic matters with enzymes. Four strains, AP45, ASP1, ASP2, and ASP4, were isolated from the coastal water of Taiwan, and respectively identified as Aurantiochytrium sp., Schizochytrium sp., Parietichytrium sp., and Botryochytrium sp. based on 18S rRNA sequences. Transcriptome datasets of these four strains at days 3-5 were generated using Next Generation Sequencing technology, and screened for enzymes with potential industrial applications. Functional annotations based on KEGG database suggest that many unigenes of all four strains were related to the pathways of industrial enzymes. Most of all four strains contained homologous genes for 15 out of the 17 targeted enzymes, and had extra- and/or intra-cellular enzymatic activities, including urease, asparaginase, lipase, glucosidase, alkaline phosphatase and protease. Complete amino sequences of the first-time identified L-asparaginase and phytase in thraustochytrids were retrieved, and respectively categorized to the Type I and BPPhy families based on phylogenetic relationships, protein structural modeling and active sites. Milligram quantities of highly purified, soluble protein of urease and L-asparaginase were successfully harvested and analyzed for recombinant enzymatic activities. These analytical results highlight the diverse enzymes for wide-range applications in thraustochytrids.
Highlights
Thraustochytrids (Kingdom Chromista: Superphylum Heterokonta: Phylum Bygira: Class Labyrinthulea: Order Thraustochytrida: Family Thraustochytriidae) are fungus-like protists categorized into at least ten genera and 40 species (Honda et al, 1999; Dellero et al, 2018a; Marchan et al, 2018; Morabito et al, 2019)
AP45, ASP1, and ASP4 were nested in the sequences of Aurantiochytrium sp., Schizochytrium sp., and Botryochytrium sp., respectively (Figure 1)
L-asparaginase and urease had only one homologous unigene identified in each strain (Table 4)
Summary
Thraustochytrids (Kingdom Chromista: Superphylum Heterokonta: Phylum Bygira: Class Labyrinthulea: Order Thraustochytrida: Family Thraustochytriidae) are fungus-like protists categorized into at least ten genera and 40 species (Honda et al, 1999; Dellero et al, 2018a; Marchan et al, 2018; Morabito et al, 2019). Several thraustochytrid strains have been successfully cultured and applied to produce high-value functional compounds. DHA plays significant roles in many aspects of animal growth and development, such as cell signaling, cell membrane fluidity, and improvement of inflammation response systems and learning (Horrocks and Yeo, 1999; Colomer et al, 2007; Aasen et al, 2016). Other promising highvalue products of thraustochytrids are carotenoids (Aki et al, 2003; Aasen et al, 2016; Park et al, 2018), including β-carotene, astaxanthin, canthaxanthin, pheonicoxanthin, and echinenone. Astaxanthin is valuable, because of its highly valued human health benefits (reviewed by Higuera-Ciapara et al, 2006). Carotenoids are often adopted in food, cosmetics and animal feed, due to their coloring and antioxidative functions (reviewed by Mezzomo and Ferreira, 2016)
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