Automated bionanocatalyst production

Abstract

The present invention provides machines, compositions and methods for producing bionanocatalysts (BNCs) comprising one or more enzymes selected from a broad spectrum of industrially and medically important enzymes. The BNCs are self-assembled and magnetically immobilized enzymes. The machines, compositions, and methods are fully scalable from bench top to industrial manufacturing volumes.

1 . A machine for the automated production of bionanocatalysts (BNC), comprising:
 a. an enzyme container;   b. a magnetic nanoparticle (MNP) container;   c. an enzyme pump;   d. an MNP pump;   e. an MNP disruptor; and   f. a BNC mixer;   
       wherein said enzyme container is operable to hold an enzyme preparation; wherein said enzyme preparation catalyzes the conversion of a diffusible substrate to a diffusible product; wherein said MNP container is operable to hold an MNP preparation; wherein said MNP pump is operable to send said MNP preparation to said MNP disruptor; wherein said MNP pump is operable to send a plurality of disrupted MNPs from said MNP disruptor to said BNC mixer; wherein said enzyme pump is operable to send an enzyme preparation to said BNC mixer; wherein said mixer is operable to mix said disrupted MNPs and enzyme preparation to form said BNCs. 
     
     
         2 . The machine of  claim 1 , wherein said MNP disruptor is a sonicator. 
     
     
         3 . The machine of  claim 2 , wherein said sonicator further comprises a sonicator coil and a sonication container, wherein said sonicator coil is operable to sonicate said MNPs within said sonication container. 
     
     
         4 . The machine of  claim 2 , wherein the sonicator is an in-line sonicator. 
     
     
         5 . The machine of  claim 2 , further comprising a cooling system operable for cooling said sonicator. 
     
     
         6 . The machine of  claim 5 , wherein said cooling system is a water cooling system. 
     
     
         7 . The machine of  claim 1 , wherein said MNP disruptor is operable to mechanically disrupt said MNPs. 
     
     
         8 . The machine of  claim 1 , wherein said MNP disruptor is operable to magnetically disrupt said MNPs. 
     
     
         9 . The machine of  claim 1 , wherein said MNP disruptor is operable to thermally disrupt said MNPs. 
     
     
         10 . The machine of  claim 1 , wherein said BNC mixer comprises a mixing tee. 
     
     
         11 . The machine of  claim 1 , wherein said BNC mixer comprises a mixing coil. 
     
     
         12 . The machine of  claim 1 , wherein said enzyme pump sends said enzyme preparation to said BNC mixer via mechanical or gravitational force. 
     
     
         13 . The machine of  claim 1 , wherein said MNP pump sends said MNP preparation to said MNP disruptor via mechanical or gravitational force. 
     
     
         14 . The machine of  claim 1 , further comprising a magnetic scaffolding container operable for mixing a magnetic scaffolding preparation with said BNCs in said scaffolding container to produce BNCs in a level 2 assembly. 
     
     
         15 . The machine of  claim 14 , wherein said magnetic scaffolding container is operable to mix said BNCS and said Scaffolding mechanically. 
     
     
         16 . The machine of  claim 14 , wherein said magnetic scaffolding container is operable to mix said BNCS and said Scaffolding magnetically. 
     
     
         17 . The machine of  claim 14 , wherein said magnetic level 2 assembly is a Magnetic Microparticle (MMP). 
     
     
         18 . The machine of  claim 1 , further comprising a templator for assembling said BNCs into a stabilizing level 2 assembly. 
     
     
         19 . The machine of  claim 18 , wherein said level 2 assembly is magnetic. 
     
     
         20 .- 32 . (canceled) 
     
     
         33 . A self-assembled bionanocatalyst (BNC) composition, comprising magnetic nanoparticles (MNPs) magnetically immobilizing an enzyme selected from the group consisting of hydrolases, hydroxylases, hydratases, transaminases, ene reductases (EREDS), imine reductases (IREDS), oxynitrilases, synthetases, and isomerases; wherein said enzyme preparation catalyzes the conversion of a diffusible substrate to a diffusible product. 
     
     
         34 . The BNC composition of  claim 33 , wherein said enzyme is at a concentration of about 5-2,000 μg per ml of total solution and said MNPs are at a concentration of about 50-20,000 μg per ml of total solution. 
     
     
         35 . The BNC composition of  claim 34 , wherein said enzyme is at a concentration of about 5-15,000 μg per ml of total solution. 
     
     
         36 . The BNC composition of  claim 35 , wherein said enzyme is at a concentration of about 5-10,000 μg per ml of total solution. 
     
     
         37 . The BNC composition of  claim 36 , wherein said enzyme is at a concentration of about 5-5,000 μg per ml of total solution. 
     
     
         38 . The BNC composition of  claim 34 , wherein said enzyme is at a concentration of about 100-20,000 μg per ml of total solution. 
     
     
         39 . The BNC composition of  claim 38 , wherein said MNPs are at a concentration of about 500-20,000 μg per ml of total solution. 
     
     
         40 . The BNC composition of  claim 39 , wherein said MNPs are at a concentration of about 1000-20,000 μg per ml of total solution. 
     
     
         41 . The BNC composition of  claim 40 , wherein said MNPs are at a concentration of about 5000-20,000 μg per ml of total solution. 
     
     
         42 . The BNC composition of  claim 41 , wherein said MNPs are at a concentration of about 10,000-20,000 μg per ml of total solution. 
     
     
         43 . The BNC composition of  claim 34 , wherein said MNPs are at a concentration of about 5000-10,000 μg per ml of total solution. 
     
     
         44 . The BNC composition of  claim 1 , further comprising a stabilizing level 2 assembly. 
     
     
         45 . The BNC composition of  claim 44 , wherein said level 2 assembly is magnetic. 
     
     
         46 . The BNC composition of  claim 45 , wherein said magnetic level 2 assembly is a Magnetic Microparticle (MMP). 
     
     
         47 . A method of using the BNC composition of  claim 33 , comprising contacting said BNC with said diffusible substrate and collecting said diffusible product. 
     
     
         48 . The BNC composition of  claim 1 , wherein said BNC composition is templated into a macro structure. 
     
     
         49 . The BNC composition of  claim 48 , wherein said macro structure is for an industrial or a medical application. 
     
     
         50 . The BNC composition of  claim 48 , wherein said macro structure comprises an MMP. 
     
     
         51 . A self-assembled bionanocatalyst (BNC) composition, comprising magnetic nanoparticles (MNPs) magnetically immobilizing a P450 oxidoreductase. 
     
     
         52 . The BNC composition of  claim 51 , further comprising a stabilizing level 2 assembly. 
     
     
         53 . The BNC composition of  claim 52 , wherein said level 2 assembly is magnetic. 
     
     
         54 . The BNC composition of  claim 53 , wherein said magnetic level 2 assembly is a Magnetic Microparticle (MMP). 
     
     
         55 . The BNC composition of  claim 51 , wherein said BNC composition is templated into a macro structure. 
     
     
         56 . The BNC composition of  claim 55 , wherein said macro structure is for a medical application. 
     
     
         57 . The BNC composition of  claim 55 , wherein said macro structure comprises an MMP. 
     
     
         58 . The BNC composition of  claim 56 , wherein said medical application is the production of a metabolite. 
     
     
         59 . The BNC composition of  claim 58 , further comprising a cofactor regenerating system. 
     
     
         60 . The BNC composition of  claim 59 , wherein said cofactor regenerating system comprises NADPH or NADP+.