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Method and cell line for production of polyketides in yeast

A technology of yeast cells and compounds, applied in biochemical equipment and methods, using vectors to introduce foreign genetic material, enzymes, etc., can solve labor-intensive and expensive problems

Pending Publication Date: 2019-12-31
海牙森生物公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Phytocannabinoid analogs are often chemically synthesized, which can be labor-intensive and expensive

Method used

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  • Method and cell line for production of polyketides in yeast
  • Method and cell line for production of polyketides in yeast
  • Method and cell line for production of polyketides in yeast

Examples

Experimental program
Comparison scheme
Effect test

Embodiment I

[0127] Yeast strain HB80 as described in Table 3 above was cultured in YNB + 2% raffinose + 2% glucose + 1.6 g / L4DO* medium. Production of methyl olivetol from raffinose and glucose was observed, suggesting direct production of methyl olivetol in yeast. Methyl olivetol was produced at a concentration of 3.259 mg / L.

Embodiment II

[0129] Yeast strain HB80A as described in Table 3 above was cultured in YNB + 2% raffinose + 2% galactose + 1.6 g / L4DO* medium. observed by DiPKS G1516D；G1518A Catalytically, both olivetol and methyl olivetol are produced from raffinose and galactose. This data indicates the direct production of olivetol and methyl olivetol in yeast excluding caproic acid.

[0130] Figure 10 Concentrations of HB80-produced methyl olivetol (“Methyl olivetol HB80”) and HB80A-produced olivetol and methyl olivetol (“Methyl olivetol HB80A” and “Olivetol HB80A” respectively) from Example 1 are shown. ")concentration. Cultures were sampled at 72 hours. HB80A mainly produces olivetol (1.4 mg olivetol per L culture compared to 0.010 mg olivetol per L culture) and produces less olivetol and olivetol in total than HB80 Olive alcohol (3.26mg / L).

Embodiment III

[0132] Yeast strain HB98 as described in Table 3 above was cultured in YNB + 2% raffinose + 2% galactose + 1.6 g / L4DO* medium. The production of methyl olivetol from raffinose and galactose catalyzed by DiPKS was observed. This data demonstrates the increased production of methyl olivetol compared to HB80 as described in Example 1, and also without the inclusion of caproic acid.

[0133] Figure 11 Concentrations of methyl olivetol produced by HB80 from Example I ("methyl olivetol HB80") and concentrations of methyl olivetol produced by HB98 from Example III ("methyl olivetol HB98") are shown. Cultures were sampled at 72 hours. HB98 produced 29.85 mg / L of olivetol, while HB80 produced only 3.26 mg of olivetol per L of culture. HB98 produced almost 10 times more methyl olivetol than HB80.

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Abstract

The present invention provides a method and a cell line for producing polyketides in yeast. The method applies, and the cell line includes, a yeast cell transformed with a polyketide synthase coding sequence. The polyketide synthase enzyme catalyzes synthesis of olivetol or methyl-olivetol, and may include Dictyostelium discoideum polyketide synthase ('DiPKS'). Wild type DiPKS produces methyl-olivetol only. DiPKS may be modified to produce olivetol only or a mixture of both olivetol and methyl-olivetol. The yeast cell may be modified to include a phosphopantethienyl transferase for increased activity of DiPKS. The yeast cell may be modified to mitigate mitochondrial acetaldehyde catabolism for increasing malonyl-CoA available for synthesizing olivetol or methyl-olivetol.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 460,526, entitled "Methods and Cell Lines for the Production of Phytocannabinoids in Yeast," filed February 17, 2017, which is hereby incorporated by reference Incorporated as a whole. technical field [0003] The present disclosure generally relates to the production of polyketide analogs in yeast. Background technique [0004] Polyketides are precursors to many valuable secondary metabolites in plants. For example, phytocannabinoids, which are produced naturally in cannabis (Cannabissativa), other plants, and some fungi, are commercially important. More than 105 phytocannabinoids are known to be biosynthesized in cannabis, or to result from the thermal or other breakdown of phytocannabinoids biosynthesized in cannabis. While the hemp plant is also a valuable source of grain, fiber and other materials, growing hemp for phy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N1/19C12N15/81C12N9/00C12P7/02C12P7/20
CPCC07C65/05C07C39/08C12N15/52C12Y203/01C12P7/22C07K14/395C12N9/1029C12N9/1288C12N9/0006C12Y602/01001C12Y101/01001C12N9/93C12Y207/08007C12N9/1085C12Y205/01C12Y205/01041
Inventor 肖哈姆·慕克吉亚历山大·詹姆斯·坎贝尔扎卡里·道格拉斯·威尔特希尔凯文·约翰·陈
Owner 海牙森生物公司