Systems and conductive structures for determining enzymatic activity and methods of formation

Abstract

Methods for determining thermophilic enzymatic activity include heating a substrate solution in a plurality of closed volumes to a predetermined reaction temperature. Without opening the closed volumes, at least one enzyme is added, substantially simultaneously, to the closed volumes. At the predetermined reaction temperature, the closed volumes are agitated and then the activity of the at least one enzyme is determined. The methods are conducive for characterizing enzymes of high-temperature reactions, with insoluble substrates, with substrates and enzymes that do not readily intermix, and with low volumes of substrate and enzyme. Systems for characterizing the enzymes are also disclosed.

1 . A system for determining enzymatic activity, the system comprising:
 a conductive structure defining:
 a plurality of wells protruding into the conductive structure from an upper surface of the conductive structure, the plurality of wells configured to receive a plurality of reaction vessels; and 
 at least one engagement feature on a sidewall of the conductive structure, the at least one engagement feature configured to engage a counterpart engagement feature to secure the conductive structure to an agitator. 
   
     
     
         2 . The system of  claim 1 , further comprising:
 an injector support structure comprising conduits configured to receive injectors and to support the injectors over a corresponding number of wells of the plurality of wells defined in the conductive structure, the injector support structure defining:
 upper openings having an upper opening width and defined in an upper surface of the injector support structure; and 
 lower openings having a lower opening width and defined in a lower surface of the injector support structure, the lower opening width being less than the upper opening width, 
 each of the conduits extending between one of the upper openings and one of the lower openings. 
   
     
     
         3 . The system of  claim 2 , wherein the injector support structure comprises a nonconductive material. 
     
     
         4 . The system of  claim 2 , wherein the injector support structure further comprises ledges, each of the ledges surrounding one of the lower openings. 
     
     
         5 . The system of  claim 2 , wherein the injector support structure further comprises extensions protruding from a supportive body, the conduits defined in the supportive body of the injector support structure, the extensions spaced from one another by a width of a row of the plurality of wells. 
     
     
         6 . The system of  claim 1 , further comprising a support structure configured to be secured to the agitator and comprising the counterpart engagement feature. 
     
     
         7 . The system of  claim 6 , wherein:
 the at least one engagement feature on the sidewall of the conductive structure comprises openings extending from a front surface of the conductive structure to a rear surface of the conductive structure; and   the counterpart engagement feature comprises threaded rods at least partially screwed into the support structure.   
     
     
         8 . The system of  claim 1 , further comprising a lid for the conductive structure, the lid comprising sidewalls extending a height of lids secured to the reaction vessels of the plurality of reaction vessels. 
     
     
         9 . The system of  claim 1 , further comprising a lid for the conductive structure, the lid defining therein a thermometer opening and at least one other opening. 
     
     
         10 . A conductive structure for supporting and heating a plurality of reaction vessels during agitation, the conductive structure comprising a block of a conductive material defining therein a plurality of wells protruding into the block from a first surface of the block and comprising at least one engagement feature protruding into the block from a sidewall surface of the block. 
     
     
         11 . The conductive structure of  claim 10 , wherein the conductive material comprises aluminum. 
     
     
         12 . The conductive structure of  claim 10 , further comprising at least one probe opening protruding into the block from at least one surface of the block. 
     
     
         13 . The conductive structure of  claim 12 , wherein the at least one probe opening comprises a thermometer opening protruding into the block from the first surface of the block. 
     
     
         14 . The conductive structure of  claim 13 , wherein the thermometer opening protrudes to a depth approximately even with a depth of the wells of the plurality of wells. 
     
     
         15 . The conductive structure of  claim 12 , wherein the at least one probe opening comprises a thermocouple opening protruding into the block from a second surface of the block, the second surface opposing the first surface. 
     
     
         16 . The conductive structure of  claim 10 , wherein the at least one engagement feature comprises at least one opening protruding horizontally into the block from the sidewall surface of the block. 
     
     
         17 . The conductive structure of  claim 10 , wherein each well of the plurality of wells defines a cylindrical and vertical sidewall and a horizontal floor. 
     
     
         18 . A method for forming a conductive structure for supporting and heating a plurality of reaction vessels during agitation, the method comprising:
 machining into a block of a conductive material from a first surface to define a plura of wells protruding into the block from the first surface of the block; and   machining into the block from a sidewall surface to define at least one engagement feature protruding into the block from the sidewall surface.   
     
     
         19 . The method of  claim 18 , wherein machining into the block from a sidewall surface to define at least one engagement feature comprises machining through the block from the sidewall surface to an opposing sidewall surface to define the at least one engagement feature extending through the block. 
     
     
         20 . The method of  claim 18 , further comprising:
 machining into the block from the first surface to define a probe opening protruding into the block; and   machining into the block from a second surface, opposing the first surface, to define another probe opening opposing the probe opening.