Structural and functional analyses of chitinolytic enzymes in the nacreous layer of
In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When sh...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Zhu, Lingxiao [verfasserIn] Shimizu, Keisuke [verfasserIn] Kintsu, Hiroyuki [verfasserIn] Negishi, Lumi [verfasserIn] Zheng, Zehua [verfasserIn] Kurumizaka, Hitoshi [verfasserIn] Sakuda, Shohei [verfasserIn] Kuriyama, Isao [verfasserIn] Atsumi, Takashi [verfasserIn] Maeyama, Kaoru [verfasserIn] Nagai, Kiyohito [verfasserIn] Kawabata, Michiyo [verfasserIn] Kohtsuka, Hisanori [verfasserIn] Miura, Toru [verfasserIn] Oka, Yoshitaka [verfasserIn] Ifuku, Shinsuke [verfasserIn] Nagata, Koji [verfasserIn] Suzuki, Michio [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2022 |
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| Schlagwörter: |
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| Übergeordnetes Werk: |
Enthalten in: Biochemical engineering journal - Amsterdam [u.a.] : Elsevier, 1998, 191 |
|---|---|
| Übergeordnetes Werk: |
volume:191 |
| DOI / URN: |
10.1016/j.bej.2022.108780 |
|---|
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| 520 | |a In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. | ||
| 650 | 4 | |a Chitin | |
| 650 | 4 | |a Chitinase | |
| 650 | 4 | |a Mineral bridge | |
| 650 | 4 | |a Nacreous layer | |
| 700 | 1 | |a Shimizu, Keisuke |e verfasserin |4 aut | |
| 700 | 1 | |a Kintsu, Hiroyuki |e verfasserin |4 aut | |
| 700 | 1 | |a Negishi, Lumi |e verfasserin |4 aut | |
| 700 | 1 | |a Zheng, Zehua |e verfasserin |4 aut | |
| 700 | 1 | |a Kurumizaka, Hitoshi |e verfasserin |4 aut | |
| 700 | 1 | |a Sakuda, Shohei |e verfasserin |4 aut | |
| 700 | 1 | |a Kuriyama, Isao |e verfasserin |4 aut | |
| 700 | 1 | |a Atsumi, Takashi |e verfasserin |4 aut | |
| 700 | 1 | |a Maeyama, Kaoru |e verfasserin |4 aut | |
| 700 | 1 | |a Nagai, Kiyohito |e verfasserin |4 aut | |
| 700 | 1 | |a Kawabata, Michiyo |e verfasserin |4 aut | |
| 700 | 1 | |a Kohtsuka, Hisanori |e verfasserin |4 aut | |
| 700 | 1 | |a Miura, Toru |e verfasserin |4 aut | |
| 700 | 1 | |a Oka, Yoshitaka |e verfasserin |4 aut | |
| 700 | 1 | |a Ifuku, Shinsuke |e verfasserin |4 aut | |
| 700 | 1 | |a Nagata, Koji |e verfasserin |4 aut | |
| 700 | 1 | |a Suzuki, Michio |e verfasserin |4 aut | |
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10.1016/j.bej.2022.108780 doi (DE-627)ELV009142630 (ELSEVIER)S1369-703X(22)00449-1 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Zhu, Lingxiao verfasserin aut Structural and functional analyses of chitinolytic enzymes in the nacreous layer of 2022 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. Chitin Chitinase Mineral bridge Nacreous layer Shimizu, Keisuke verfasserin aut Kintsu, Hiroyuki verfasserin aut Negishi, Lumi verfasserin aut Zheng, Zehua verfasserin aut Kurumizaka, Hitoshi verfasserin aut Sakuda, Shohei verfasserin aut Kuriyama, Isao verfasserin aut Atsumi, Takashi verfasserin aut Maeyama, Kaoru verfasserin aut Nagai, Kiyohito verfasserin aut Kawabata, Michiyo verfasserin aut Kohtsuka, Hisanori verfasserin aut Miura, Toru verfasserin aut Oka, Yoshitaka verfasserin aut Ifuku, Shinsuke verfasserin aut Nagata, Koji verfasserin aut Suzuki, Michio verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 191 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:191 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 191 |
| spelling |
10.1016/j.bej.2022.108780 doi (DE-627)ELV009142630 (ELSEVIER)S1369-703X(22)00449-1 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Zhu, Lingxiao verfasserin aut Structural and functional analyses of chitinolytic enzymes in the nacreous layer of 2022 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. Chitin Chitinase Mineral bridge Nacreous layer Shimizu, Keisuke verfasserin aut Kintsu, Hiroyuki verfasserin aut Negishi, Lumi verfasserin aut Zheng, Zehua verfasserin aut Kurumizaka, Hitoshi verfasserin aut Sakuda, Shohei verfasserin aut Kuriyama, Isao verfasserin aut Atsumi, Takashi verfasserin aut Maeyama, Kaoru verfasserin aut Nagai, Kiyohito verfasserin aut Kawabata, Michiyo verfasserin aut Kohtsuka, Hisanori verfasserin aut Miura, Toru verfasserin aut Oka, Yoshitaka verfasserin aut Ifuku, Shinsuke verfasserin aut Nagata, Koji verfasserin aut Suzuki, Michio verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 191 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:191 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 191 |
| allfields_unstemmed |
10.1016/j.bej.2022.108780 doi (DE-627)ELV009142630 (ELSEVIER)S1369-703X(22)00449-1 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Zhu, Lingxiao verfasserin aut Structural and functional analyses of chitinolytic enzymes in the nacreous layer of 2022 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. Chitin Chitinase Mineral bridge Nacreous layer Shimizu, Keisuke verfasserin aut Kintsu, Hiroyuki verfasserin aut Negishi, Lumi verfasserin aut Zheng, Zehua verfasserin aut Kurumizaka, Hitoshi verfasserin aut Sakuda, Shohei verfasserin aut Kuriyama, Isao verfasserin aut Atsumi, Takashi verfasserin aut Maeyama, Kaoru verfasserin aut Nagai, Kiyohito verfasserin aut Kawabata, Michiyo verfasserin aut Kohtsuka, Hisanori verfasserin aut Miura, Toru verfasserin aut Oka, Yoshitaka verfasserin aut Ifuku, Shinsuke verfasserin aut Nagata, Koji verfasserin aut Suzuki, Michio verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 191 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:191 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 191 |
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10.1016/j.bej.2022.108780 doi (DE-627)ELV009142630 (ELSEVIER)S1369-703X(22)00449-1 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Zhu, Lingxiao verfasserin aut Structural and functional analyses of chitinolytic enzymes in the nacreous layer of 2022 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. Chitin Chitinase Mineral bridge Nacreous layer Shimizu, Keisuke verfasserin aut Kintsu, Hiroyuki verfasserin aut Negishi, Lumi verfasserin aut Zheng, Zehua verfasserin aut Kurumizaka, Hitoshi verfasserin aut Sakuda, Shohei verfasserin aut Kuriyama, Isao verfasserin aut Atsumi, Takashi verfasserin aut Maeyama, Kaoru verfasserin aut Nagai, Kiyohito verfasserin aut Kawabata, Michiyo verfasserin aut Kohtsuka, Hisanori verfasserin aut Miura, Toru verfasserin aut Oka, Yoshitaka verfasserin aut Ifuku, Shinsuke verfasserin aut Nagata, Koji verfasserin aut Suzuki, Michio verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 191 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:191 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 191 |
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10.1016/j.bej.2022.108780 doi (DE-627)ELV009142630 (ELSEVIER)S1369-703X(22)00449-1 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Zhu, Lingxiao verfasserin aut Structural and functional analyses of chitinolytic enzymes in the nacreous layer of 2022 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. Chitin Chitinase Mineral bridge Nacreous layer Shimizu, Keisuke verfasserin aut Kintsu, Hiroyuki verfasserin aut Negishi, Lumi verfasserin aut Zheng, Zehua verfasserin aut Kurumizaka, Hitoshi verfasserin aut Sakuda, Shohei verfasserin aut Kuriyama, Isao verfasserin aut Atsumi, Takashi verfasserin aut Maeyama, Kaoru verfasserin aut Nagai, Kiyohito verfasserin aut Kawabata, Michiyo verfasserin aut Kohtsuka, Hisanori verfasserin aut Miura, Toru verfasserin aut Oka, Yoshitaka verfasserin aut Ifuku, Shinsuke verfasserin aut Nagata, Koji verfasserin aut Suzuki, Michio verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 191 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:191 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 191 |
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Enthalten in Biochemical engineering journal 191 volume:191 |
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Chitin Chitinase Mineral bridge Nacreous layer |
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Zhu, Lingxiao @@aut@@ Shimizu, Keisuke @@aut@@ Kintsu, Hiroyuki @@aut@@ Negishi, Lumi @@aut@@ Zheng, Zehua @@aut@@ Kurumizaka, Hitoshi @@aut@@ Sakuda, Shohei @@aut@@ Kuriyama, Isao @@aut@@ Atsumi, Takashi @@aut@@ Maeyama, Kaoru @@aut@@ Nagai, Kiyohito @@aut@@ Kawabata, Michiyo @@aut@@ Kohtsuka, Hisanori @@aut@@ Miura, Toru @@aut@@ Oka, Yoshitaka @@aut@@ Ifuku, Shinsuke @@aut@@ Nagata, Koji @@aut@@ Suzuki, Michio @@aut@@ |
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2022-01-01T00:00:00Z |
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Zhu, Lingxiao |
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660 540 DE-600 58.30 bkl 58.00 bkl Structural and functional analyses of chitinolytic enzymes in the nacreous layer of Chitin Chitinase Mineral bridge Nacreous layer |
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Zhu, Lingxiao Shimizu, Keisuke Kintsu, Hiroyuki Negishi, Lumi Zheng, Zehua Kurumizaka, Hitoshi Sakuda, Shohei Kuriyama, Isao Atsumi, Takashi Maeyama, Kaoru Nagai, Kiyohito Kawabata, Michiyo Kohtsuka, Hisanori Miura, Toru Oka, Yoshitaka Ifuku, Shinsuke Nagata, Koji Suzuki, Michio |
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structural and functional analyses of chitinolytic enzymes in the nacreous layer of |
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Structural and functional analyses of chitinolytic enzymes in the nacreous layer of |
| abstract |
In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. |
| abstractGer |
In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. |
| abstract_unstemmed |
In this study, chitinolytic activity was confirmed in the protein extracted from the inner nacreous layer of Pinctada fucata shells. Allosamidin, a specific inhibitor of family 18 chitinase, was injected into pearl oysters, and an abnormal organic membrane was observed in the nacreous layer. When shell-notched individuals were injected with allosamidin, the organic fibers in the regenerated site were thicker than those without allosamidin treatment. To identify chitinases in the nacreous layer, proteins extracted from the nacreous layer were analyzed using LC-MS/MS. Two chitinases (PfCN1 and PfCN2) were identified, and their different spatial expression patterns were observed in mantle tissue. To understand the function of chitinolytic enzymes in nacre formation, we designed an in vivo gene knockdown experiment using PfCN1 and PfCN2 dsRNA, and in vitro calcium carbonate crystallization experiments using chitinolytic enzymes. In the knockdown experiment, abnormal aragonite tablets and organic films were observed in the nacreous layer. In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. These results showed the novel function of chitinolytic enzymes that will give a comprehension of nacre formation. |
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Shimizu, Keisuke Kintsu, Hiroyuki Negishi, Lumi Zheng, Zehua Kurumizaka, Hitoshi Sakuda, Shohei Kuriyama, Isao Atsumi, Takashi Maeyama, Kaoru Nagai, Kiyohito Kawabata, Michiyo Kohtsuka, Hisanori Miura, Toru Oka, Yoshitaka Ifuku, Shinsuke Nagata, Koji Suzuki, Michio |
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In the crystallization experiment, we found that the thickness of the chitin film decreased and the space between the chitin fibers increased when calcium carbonate was formed on the chitin film on the glass substrate with chitin-degrading enzyme treatment. These reactions induce the growth of calcium carbonate crystals through the chitin film. The penetration of chitin film by calcium carbonate is similar to that of mineral bridges that connect the nacre tablets vertically. 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7.400487 |