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公开(公告)号：US20210032635A1
公开(公告)日：2021-02-04
申请号：US16651043
申请日：2018-09-28
Applicant: VICTORIA LINK LIMITED
Abstract: A modular and hierarchical DNA assembly platform for synthetic biology is described. This enabling technology, termed MIDAS (for Modular Idempotent DNA Assembly System), can precisely assemble multiple DNA fragments in a single reaction using a standardised assembly design. It can be used to build genes from libraries of sequence-verified, reusable parts and to assemble multiple genes in a single vector. We describe the design and use of MIDAS, and its application in the reconstruction of the metabolic pathway for production of paspaline, a key intermediate in the biosynthesis of a range of indole diterpenes—a class of economically important secondary metabolites produced by several species of filamentous fungi.
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Patent Agency Ranking
公开(公告)号：US20200299700A1
公开(公告)日：2020-09-24
申请号：US16651065
申请日：2018-09-28
Applicant: VICTORIA LINK LIMITED
Inventor： Matthew Joseph Nicholson , Sarah Adeline Kessans , Emily Jane Parker , Leyla Yolanda Bustamante Rodriguez , David Barry Scott , Kyle Cornelius Van de Bittner , Craig John Van Dolleweerd
Abstract: Nodulisporic acids (NAs) comprise a group of indole diterpenes known for their potent insecticidal activities; however, biosynthesis of NAs by its natural producer, Hypoxylon pulicicidum (Nodulisporium sp.) is exceptionally difficult to achieve. The identification of genes responsible for NA production could enable biosynthetic pathway optimization to provide access to NAs for commercial applications. Obtaining useful quantities of NAs using published fermentations methods is challenging, making gene knockout studies an undesirable method to confirm gene function. Alternatively, heterologous gene expression of H. pulicicidum genes in a more robust host species like Penicillium paxilli provides a way to rapidly identify the function of genes that play a role in NA biosynthesis. In this work, we identified the function of four secondary-metabolic genes necessary for the biosynthesis of nodulisporic acid F (NAF) and reconstituted these genes in the genome of P. paxilli to enable heterologous production of NAF in this fungus.
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公开(公告)号：US12065650B2
公开(公告)日：2024-08-20
申请号：US17816630
申请日：2022-08-01
Applicant: VICTORIA LINK LIMITED
Inventor： Matthew Joseph Nicholson , Sarah Adeline Kessans , Emily Jane Parker , Leyla Yolanda Bustamante Rodriguez , David Barry Scott , Kyle Cornelius Van de Bittner , Craig John Van Dolleweerd
CPC classification number: C12N15/52 , A01N63/30 , A01N63/50 , C12N15/80 , C12P17/18 , A01N63/50 , A01N63/30
Abstract: Nodulisporic acids (NAs) comprise a group of indole diterpenes known for their potent insecticidal activities; however, biosynthesis of NAs by its natural producer, Hypoxylon pulicicidum (Nodulisporium sp.) is exceptionally difficult to achieve. The identification of genes responsible for NA production could enable biosynthetic pathway optimization to provide access to NAs for commercial applications. Obtaining useful quantities of NAs using published fermentations methods is challenging, making gene knockout studies an undesirable method to confirm gene function. Alternatively, heterologous gene expression of H. pulicicidum genes in a more robust host species like Penicillium paxilli provides a way to rapidly identify the function of genes that play a role in NA biosynthesis. In this work, we identified the function of four secondary-metabolic genes necessary for the biosynthesis of nodulisporic acid F (NAF) and reconstituted these genes in the genome of P. paxilli to enable heterologous production of NAF in this fungus.
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公开(公告)号：US20230365976A1
公开(公告)日：2023-11-16
申请号：US17816630
申请日：2022-08-01
Applicant: VICTORIA LINK LIMITED
Inventor： Matthew Joseph Nicholson , Sarah Adeline Kessans , Emily Jane Parker , Leyla Yolanda Bustamante Rodriguez , David Barry Scott , Kyle Cornelius Van de Bittner , Craig John Van Dolleweerd
Abstract: Nodulisporic acids (NAs) comprise a group of indole diterpenes known for their potent insecticidal activities; however, biosynthesis of NAs by its natural producer, Hypoxylon pulicicidum (Nodulisporium sp.) is exceptionally difficult to achieve. The identification of genes responsible for NA production could enable biosynthetic pathway optimization to provide access to NAs for commercial applications. Obtaining useful quantities of NAs using published fermentations methods is challenging, making gene knockout studies an undesirable method to confirm gene function. Alternatively, heterologous gene expression of H. pulicicidum genes in a more robust host species like Penicillium paxilli provides a way to rapidly identify the function of genes that play a role in NA biosynthesis. In this work, we identified the function of four secondary-metabolic genes necessary for the biosynthesis of nodulisporic acid F (NAF) and reconstituted these genes in the genome of P. paxilli to enable heterologous production of NAF in this fungus.
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公开(公告)号：US11655476B2
公开(公告)日：2023-05-23
申请号：US16651043
申请日：2018-09-28
Applicant: VICTORIA LINK LIMITED
CPC classification number: C12N15/64 , C12N15/52 , C12N15/66 , C12N15/70 , C12P17/188 , C12N2510/02 , C12N2800/204 , C12N2800/40 , C12N2800/50
Abstract: A modular and hierarchical DNA assembly platform for synthetic biology is described. This enabling technology, termed MIDAS (for Modular Idempotent DNA Assembly System), can precisely assemble multiple DNA fragments in a single reaction using a standardised assembly design. It can be used to build genes from libraries of sequence-verified, reusable parts and to assemble multiple genes in a single vector. We describe the design and use of MIDAS, and its application in the reconstruction of the metabolic pathway for production of paspaline, a key intermediate in the biosynthesis of a range of indole diterpenes—a class of economically important secondary metabolites produced by several species of filamentous fungi.
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公开(公告)号：US11453882B2
公开(公告)日：2022-09-27
申请号：US16651065
申请日：2018-09-28
Applicant: VICTORIA LINK LIMITED
Inventor： Matthew Joseph Nicholson , Sarah Adeline Kessans , Emily Jane Parker , Leyla Yolanda Bustamante Rodriguez , David Barry Scott , Kyle Cornelius Van de Bittner , Craig John Van Dolleweerd
Abstract: Nodulisporic acids (NAs) comprise a group of indole diterpenes known for their potent insecticidal activities; however, biosynthesis of NAs by its natural producer, Hypoxylon pulicicidum (Nodulisporium sp.) is exceptionally difficult to achieve. The identification of genes responsible for NA production could enable biosynthetic pathway optimization to provide access to NAs for commercial applications. Obtaining useful quantities of NAs using published fermentations methods is challenging, making gene knockout studies an undesirable method to confirm gene function. Alternatively, heterologous gene expression of H. pulicicidum genes in a more robust host species like Penicillium paxilli provides a way to rapidly identify the function of genes that play a role in NA biosynthesis. In this work, we identified the function of four secondary-metabolic genes necessary for the biosynthesis of nodulisporic acid F (NAF) and reconstituted these genes in the genome of P. paxilli to enable heterologous production of NAF in this fungus.
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