In vitro metabolic engineering on microscale devices
First Claim
1. An apparatus for conducting a plurality of microscale reactions, comprisinga. a solid support comprising a first and a second reaction site, wherein each site comprises a distinct immobilized biocatalyst;
- b. a micro-directing element to direct an intermediate composition from the first site to the second site,wherein the biocatalyst at the first reaction site and the biocatalyst at the second reaction site are each a functional catalytic unit from sequential steps in a natural biosynthetic pathway, the apparatus further comprising a micro-directing element to direct a composition from a source to at least one said reaction site, andwherein1) the apparatus comprises n additional reaction sites;
2) n is an integer from 1 to 50;
3) each reaction site comprises a distinct immobilized biocatalyst; and
4) the apparatus additionally comprises one or more micro-directing elements to sequentially direct a composition from the second reaction site to each of the n additional reaction sites.
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Accused Products
Abstract
Disclosed herein is a microfluidics device that can be used to prepare natural products and their analogs. The device comprises the enzymes of a biosynthetic pathway immobilized thereon and a means for sequentially directing a starting material and each ensuing reaction product to the enzymes of the biosynthetic pathway in the order corresponding to the steps of the biosynthetic pathway. The device can thus be used to prepare the natural product using the natural starting material of the biosynthetic pathway or analogs of the natural product using an unnatural starting material. Alternatively, artificial pathways can be created by immobilizing an appropriate selection of enzymes on the device in an order whereby each subsequent enzyme can catalyze a reaction with the product of the prior enzyme. Novel chemical entities can be prepared from these artificial pathways.
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Citations
13 Claims
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1. An apparatus for conducting a plurality of microscale reactions, comprising
a. a solid support comprising a first and a second reaction site, wherein each site comprises a distinct immobilized biocatalyst; -
b. a micro-directing element to direct an intermediate composition from the first site to the second site, wherein the biocatalyst at the first reaction site and the biocatalyst at the second reaction site are each a functional catalytic unit from sequential steps in a natural biosynthetic pathway, the apparatus further comprising a micro-directing element to direct a composition from a source to at least one said reaction site, and wherein 1) the apparatus comprises n additional reaction sites; 2) n is an integer from 1 to 50; 3) each reaction site comprises a distinct immobilized biocatalyst; and 4) the apparatus additionally comprises one or more micro-directing elements to sequentially direct a composition from the second reaction site to each of the n additional reaction sites. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A microfluidics chip, comprising
a. a solid support comprising a first and second reaction site, wherein i. the first and second reaction sites each comprise a distinct biocatalyst that is a functional catalytic unit of a natural biosynthetic pathway; -
ii. each said biocatalyst is immobilized by a polyacrylate substituted with sucrose, lactose, mannose, an alkylmannoside, glucose, an alkylglucoside, galactose, an alkylgalactoside, fructose, an alkylfructoside, or trehalose; b. microfluidics elements to direct a substrate composition from a source to the first reactive composition, and microfluidics elements to direct an intermediate composition from the first reaction site to the second reaction site, wherein the biocatalysts at the first and second sites are from sequential steps in the natural biosynthetic pathway.
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Specification