Acaudina leucoprocta is an edible sea cucumber of economic interest that is widely distributed in China. Little information is available concerning the molecular genetics of this species although such knowledge would ...Acaudina leucoprocta is an edible sea cucumber of economic interest that is widely distributed in China. Little information is available concerning the molecular genetics of this species although such knowledge would contribute to a better understanding of the optimal conditions for its aquaculture and its mechanisms of defense against disease. Therefore, we constructed a c DNA library and, based on bioinformatics analysis of the sequences, the functions of 75% of the c DNAs were identifi ed, including those involved in cell structure, energy metabolism, mitochondrial function, and signal transduction pathways. Approximately 25% of genes in the library were unmatched. The gene for A. leucoprocta ferritin was also cloned. The predicted amino-acid sequence of ferritin displayed signifi cant homology with other sea-cucumber counterparts but indicated that it was a new member of the ferritin family. Semiquantitative real-time RT-PCR indicated the highest levels of ferritin m RNA expression in the intestine. A polyclonal antibody of ferritin was also produced. These data provide a set of molecular tools essential for further studies of the functions of ferritin protein in A. leucoprocta.展开更多
The sea potato Acaudina spp.is a species of sea cucumber,belonging to the class Holothuroidea under Phylum Echino-dermata.It becomes blooming recently in the East Sea of China,causing serious ecological problems.Howev...The sea potato Acaudina spp.is a species of sea cucumber,belonging to the class Holothuroidea under Phylum Echino-dermata.It becomes blooming recently in the East Sea of China,causing serious ecological problems.However,until now there are no molecular data for its larval identification and population genetic analysis.In this study,we firstly screened a mtDNA fragment and demonstrated that it was the species-specific molecular marker for the identification of Acaudina spp.We then developed a quantitative polymerase chain reaction(PCR)method to evaluate the larval density of Acaudina spp.based on this molecular probe.Utilizing this method,we examined 116 plankton samples collected in four seasons from 13 stations along the coastal region in Fu-jian province,China.The results showed that the high larval density was presented at stations 1,2,and 3,which were near a quay in the coast.The larval density increased from April and reached the highest value in June and July,suggesting temperature might be the main environmental factor on the effects of its population distribution and density.Our work provides an important molecular tool for species identification and risk evaluation of a potentially invasive species.展开更多
Protein hydrolysates from Acaudina leucoprocta(ALPs)exert a protective effect in alcoholic liver injury mice through the activation of alcohol dehydrogenase(ADH).However,the amino acid sequences and interaction mechan...Protein hydrolysates from Acaudina leucoprocta(ALPs)exert a protective effect in alcoholic liver injury mice through the activation of alcohol dehydrogenase(ADH).However,the amino acid sequences and interaction mechanisms of peptides in ALPs against ADH is not identified.In the present study,123 peptides were separated and identified from ALPs by volume exclusion chromatography,reversed-phase chromatography and liquid chromatography tandem mass spectrometry(LC-MS/MS).Computer-assisted screening yielded 29 peptides that were predicted to be non-toxic and non-allergenic for use.Five peptides including PPWQWDK(PK-7),YPLPK(YK-5),SLDWRAK(SK-7),GGLHLPK(GK-7)and MRLK(MK-4)were selected by amphiphilicity and spatial resistance,and synthesized for the evaluation of ADH activation rate.GK-7 showed significantly higher ADH activation than the other peptides and the fluorescence quenching data reveal a dynamic quenching constant(Kq)of 1.296×10^(10) M^(-1) s^(-1) for GK-7 with ADH,suggesting the presence of conformational changes in both.Infrared spectroscopy and circular dichroism spectroscopy further indicate that GK-7 induces the formation of stable complexes of ADH in a β-folded form.Molecular docking simulations reveal that GK-7 binds to the active center of ADH through hydrogen bonding and hydrophobic interactions.These results suggest that GK-7 stabilizes and activates ADH and improves its catalytic efficiency.In conclusion,the findings further demonstrate that GK-7 can promote alcohol metabolism and prevent alcoholic liver injury,and have potential for development of novel functional foods or dietary supplements.展开更多
基金Supported by the Zhejiang Science and Technology Department(No.2009C03017-4)the Major Project of the Ningbo Science Bureau(No.2008C50027)the K.C.Wong Magna Fund at Ningbo University
文摘Acaudina leucoprocta is an edible sea cucumber of economic interest that is widely distributed in China. Little information is available concerning the molecular genetics of this species although such knowledge would contribute to a better understanding of the optimal conditions for its aquaculture and its mechanisms of defense against disease. Therefore, we constructed a c DNA library and, based on bioinformatics analysis of the sequences, the functions of 75% of the c DNAs were identifi ed, including those involved in cell structure, energy metabolism, mitochondrial function, and signal transduction pathways. Approximately 25% of genes in the library were unmatched. The gene for A. leucoprocta ferritin was also cloned. The predicted amino-acid sequence of ferritin displayed signifi cant homology with other sea-cucumber counterparts but indicated that it was a new member of the ferritin family. Semiquantitative real-time RT-PCR indicated the highest levels of ferritin m RNA expression in the intestine. A polyclonal antibody of ferritin was also produced. These data provide a set of molecular tools essential for further studies of the functions of ferritin protein in A. leucoprocta.
基金This work was supported by the National Key Research and Development Program of China(No.2018YFD0900705)the Key Laboratory of Integrated Marine Monitor-ing and Applied Technologies for Harmful Algal Blooms Funds(No.MATHAB201706)the Fundamental Re-search Funds for the Central Universities(No.201822016).
文摘The sea potato Acaudina spp.is a species of sea cucumber,belonging to the class Holothuroidea under Phylum Echino-dermata.It becomes blooming recently in the East Sea of China,causing serious ecological problems.However,until now there are no molecular data for its larval identification and population genetic analysis.In this study,we firstly screened a mtDNA fragment and demonstrated that it was the species-specific molecular marker for the identification of Acaudina spp.We then developed a quantitative polymerase chain reaction(PCR)method to evaluate the larval density of Acaudina spp.based on this molecular probe.Utilizing this method,we examined 116 plankton samples collected in four seasons from 13 stations along the coastal region in Fu-jian province,China.The results showed that the high larval density was presented at stations 1,2,and 3,which were near a quay in the coast.The larval density increased from April and reached the highest value in June and July,suggesting temperature might be the main environmental factor on the effects of its population distribution and density.Our work provides an important molecular tool for species identification and risk evaluation of a potentially invasive species.
基金supported by the Natural Science Foundation of Zhejiang Province(No.LTGY24D060002)National Natural Science Foundation of China(No.82073764)Science and Technology Plan Project of Zhoushan Putuo District of China(No.2021JH105).
文摘Protein hydrolysates from Acaudina leucoprocta(ALPs)exert a protective effect in alcoholic liver injury mice through the activation of alcohol dehydrogenase(ADH).However,the amino acid sequences and interaction mechanisms of peptides in ALPs against ADH is not identified.In the present study,123 peptides were separated and identified from ALPs by volume exclusion chromatography,reversed-phase chromatography and liquid chromatography tandem mass spectrometry(LC-MS/MS).Computer-assisted screening yielded 29 peptides that were predicted to be non-toxic and non-allergenic for use.Five peptides including PPWQWDK(PK-7),YPLPK(YK-5),SLDWRAK(SK-7),GGLHLPK(GK-7)and MRLK(MK-4)were selected by amphiphilicity and spatial resistance,and synthesized for the evaluation of ADH activation rate.GK-7 showed significantly higher ADH activation than the other peptides and the fluorescence quenching data reveal a dynamic quenching constant(Kq)of 1.296×10^(10) M^(-1) s^(-1) for GK-7 with ADH,suggesting the presence of conformational changes in both.Infrared spectroscopy and circular dichroism spectroscopy further indicate that GK-7 induces the formation of stable complexes of ADH in a β-folded form.Molecular docking simulations reveal that GK-7 binds to the active center of ADH through hydrogen bonding and hydrophobic interactions.These results suggest that GK-7 stabilizes and activates ADH and improves its catalytic efficiency.In conclusion,the findings further demonstrate that GK-7 can promote alcohol metabolism and prevent alcoholic liver injury,and have potential for development of novel functional foods or dietary supplements.
