Reconstitution of
Horseshoe crab Factor G is a heterodimeric serine protease zymogen that is activated by (1[right arrow]3)-[beta]-D-glucans (BDG) from fungal cell walls. This reaction is used in diagnostic agents for deep-seated mycosis. At present, functional analysis using Factor G from Tachypleus tridentatus has...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2023-12, Vol.107 (24), p.7463 |
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| container_title | Applied microbiology and biotechnology |
| container_volume | 107 |
| creator | Yamamoto, Yotaro Kajiura, Hiroyuki Fukuchi, Hiroki Nishibu, Takahiro Fujiyama, Kazuhito Kitagawa, Takeshi |
| description | Horseshoe crab Factor G is a heterodimeric serine protease zymogen that is activated by (1[right arrow]3)-[beta]-D-glucans (BDG) from fungal cell walls. This reaction is used in diagnostic agents for deep-seated mycosis. At present, functional analysis using Factor G from Tachypleus tridentatus has been performed, and genetic information has been published, but reconstitution using recombinant proteins has not yet been achieved. In this study, we cloned the genes for Factor G [alpha] and [beta] from Limulus polyphemus; two gene sequences were obtained for Factor G [alpha] and seven for [beta]. The obtained L. polyphemus Factor G [alpha] was used to specifically remove BDG from the culture medium for eliminating the activator BDG. The optimal combination for each sequence was examined with BDG removal medium, and a combination was found that featured BDG-dependent activity. These results indicate that a BDG assay system using recombinant Factor G is feasible in reconstitution. This research will support future reagent development that does not require natural horseshoe crab resources. |
| doi_str_mv | 10.1007/s00253-023-12808-6 |
| format | Article |
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This reaction is used in diagnostic agents for deep-seated mycosis. At present, functional analysis using Factor G from Tachypleus tridentatus has been performed, and genetic information has been published, but reconstitution using recombinant proteins has not yet been achieved. In this study, we cloned the genes for Factor G [alpha] and [beta] from Limulus polyphemus; two gene sequences were obtained for Factor G [alpha] and seven for [beta]. The obtained L. polyphemus Factor G [alpha] was used to specifically remove BDG from the culture medium for eliminating the activator BDG. The optimal combination for each sequence was examined with BDG removal medium, and a combination was found that featured BDG-dependent activity. These results indicate that a BDG assay system using recombinant Factor G is feasible in reconstitution. This research will support future reagent development that does not require natural horseshoe crab resources.</description><identifier>ISSN: 0175-7598</identifier><identifier>DOI: 10.1007/s00253-023-12808-6</identifier><language>eng</language><publisher>Springer</publisher><subject>Analysis ; DNA sequencing ; Glucans ; Identification and classification ; Methods ; Nucleotide sequencing ; Properties ; Proteases ; Zymogens</subject><ispartof>Applied microbiology and biotechnology, 2023-12, Vol.107 (24), p.7463</ispartof><rights>COPYRIGHT 2023 Springer</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids></links><search><creatorcontrib>Yamamoto, Yotaro</creatorcontrib><creatorcontrib>Kajiura, Hiroyuki</creatorcontrib><creatorcontrib>Fukuchi, Hiroki</creatorcontrib><creatorcontrib>Nishibu, Takahiro</creatorcontrib><creatorcontrib>Fujiyama, Kazuhito</creatorcontrib><creatorcontrib>Kitagawa, Takeshi</creatorcontrib><title>Reconstitution of</title><title>Applied microbiology and biotechnology</title><description>Horseshoe crab Factor G is a heterodimeric serine protease zymogen that is activated by (1[right arrow]3)-[beta]-D-glucans (BDG) from fungal cell walls. This reaction is used in diagnostic agents for deep-seated mycosis. At present, functional analysis using Factor G from Tachypleus tridentatus has been performed, and genetic information has been published, but reconstitution using recombinant proteins has not yet been achieved. In this study, we cloned the genes for Factor G [alpha] and [beta] from Limulus polyphemus; two gene sequences were obtained for Factor G [alpha] and seven for [beta]. The obtained L. polyphemus Factor G [alpha] was used to specifically remove BDG from the culture medium for eliminating the activator BDG. The optimal combination for each sequence was examined with BDG removal medium, and a combination was found that featured BDG-dependent activity. These results indicate that a BDG assay system using recombinant Factor G is feasible in reconstitution. This research will support future reagent development that does not require natural horseshoe crab resources.</description><subject>Analysis</subject><subject>DNA sequencing</subject><subject>Glucans</subject><subject>Identification and classification</subject><subject>Methods</subject><subject>Nucleotide sequencing</subject><subject>Properties</subject><subject>Proteases</subject><subject>Zymogens</subject><issn>0175-7598</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><recordid>eNpjYJAyNNAzNDAw1y82MDAyNdY1MDLWNTSyMLDQNWNh4DQwNDfVNTe1tOBg4CouzjIwAEqZmXEyCAalJufnFZdklpSWZObnKeSn8TCwpiXmFKfyQmluBk031xBnD930xJzU-Mw8oPKS1IqS9MTS4uJ4z-CgeEdzc2MjCxMjEwtjUtQCAFyLMMA</recordid><startdate>20231201</startdate><enddate>20231201</enddate><creator>Yamamoto, Yotaro</creator><creator>Kajiura, Hiroyuki</creator><creator>Fukuchi, Hiroki</creator><creator>Nishibu, Takahiro</creator><creator>Fujiyama, Kazuhito</creator><creator>Kitagawa, Takeshi</creator><general>Springer</general><scope>ISR</scope></search><sort><creationdate>20231201</creationdate><title>Reconstitution of</title><author>Yamamoto, Yotaro ; Kajiura, Hiroyuki ; Fukuchi, Hiroki ; Nishibu, Takahiro ; Fujiyama, Kazuhito ; Kitagawa, Takeshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-gale_incontextgauss_ISR_A7732842483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Analysis</topic><topic>DNA sequencing</topic><topic>Glucans</topic><topic>Identification and classification</topic><topic>Methods</topic><topic>Nucleotide sequencing</topic><topic>Properties</topic><topic>Proteases</topic><topic>Zymogens</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yamamoto, Yotaro</creatorcontrib><creatorcontrib>Kajiura, Hiroyuki</creatorcontrib><creatorcontrib>Fukuchi, Hiroki</creatorcontrib><creatorcontrib>Nishibu, Takahiro</creatorcontrib><creatorcontrib>Fujiyama, Kazuhito</creatorcontrib><creatorcontrib>Kitagawa, Takeshi</creatorcontrib><collection>Gale In Context: Science</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yamamoto, Yotaro</au><au>Kajiura, Hiroyuki</au><au>Fukuchi, Hiroki</au><au>Nishibu, Takahiro</au><au>Fujiyama, Kazuhito</au><au>Kitagawa, Takeshi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Reconstitution of</atitle><jtitle>Applied microbiology and biotechnology</jtitle><date>2023-12-01</date><risdate>2023</risdate><volume>107</volume><issue>24</issue><spage>7463</spage><pages>7463-</pages><issn>0175-7598</issn><abstract>Horseshoe crab Factor G is a heterodimeric serine protease zymogen that is activated by (1[right arrow]3)-[beta]-D-glucans (BDG) from fungal cell walls. This reaction is used in diagnostic agents for deep-seated mycosis. At present, functional analysis using Factor G from Tachypleus tridentatus has been performed, and genetic information has been published, but reconstitution using recombinant proteins has not yet been achieved. In this study, we cloned the genes for Factor G [alpha] and [beta] from Limulus polyphemus; two gene sequences were obtained for Factor G [alpha] and seven for [beta]. The obtained L. polyphemus Factor G [alpha] was used to specifically remove BDG from the culture medium for eliminating the activator BDG. The optimal combination for each sequence was examined with BDG removal medium, and a combination was found that featured BDG-dependent activity. These results indicate that a BDG assay system using recombinant Factor G is feasible in reconstitution. This research will support future reagent development that does not require natural horseshoe crab resources.</abstract><pub>Springer</pub><doi>10.1007/s00253-023-12808-6</doi><tpages>11</tpages></addata></record> |
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| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 2023-12, Vol.107 (24), p.7463 |
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| language | eng |
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| source | Springer Nature - Complete Springer Journals |
| subjects | Analysis DNA sequencing Glucans Identification and classification Methods Nucleotide sequencing Properties Proteases Zymogens |
| title | Reconstitution of |
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