Screening of microorganisms for bioremediation

• US 5,756,304 A
• Filed: 07/14/1995
• Issued: 05/26/1998
• Est. Priority Date: 07/14/1995
• Status: Expired due to Term

• Current Assignee: Molecular Solutions Co.
• Original Assignee: Molecular Solutions Co.
• Inventors: Jovanovich, Stevan B.
• Primary Examiner(s): LEARY, LOUISE N
• Application Number:
  US08/502,050
• Time in Patent Office: 1,047 Days
• Field of Search: 435/34, 435/262, 435/4, 435/287, 435/252, 435/821, 435/254.1, 435/800, 435/839, 435/832, 436/43, 436/139, 422/50, 422/68.1, 588/251, 588/900
• US Class Current: 435/34
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: G01N 35/0099 comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: Y10S 435/80 Elimination or reduction of contamination by undersired ferments, e.g. aseptic cultivation
• CPC Class Codes: Y10S 435/821 Microorganisms used in the destruction of hazardous or toxic waste
• CPC Class Codes: Y10S 435/832 Bacillus
• CPC Class Codes: Y10S 435/839 Bacillus subtilis
• CPC Class Codes: Y10S 588/90 Apparatus
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/21 Hydrocarbon

Abstract

The present invention is directed to the application of robotics to screen and optimize microorganisms for their bioremediation capabilities. In particular, the present invention provides methods to screen for the ability of microorganisms to metabolize particular compounds of interest in bioremediation applications. The present invention also provides a method for discovery of microorganisms useful for bioremediation and biomining, as well as other applications where microbial metabolism is useful for catalyzing chemical biotransformations.

Claims

1. An automated method for screening microorganisms capable of detoxifying a compound, comprising the steps of:

a. providing;

i. a plurality of samples in a pair of final holders having separate vessels, each final holder separating the samples into said vessels, said samples suspected of containing microorganisms,

ii. a chemical compound, and

iii. an automated workstation;

b. exposing said samples to said chemical compound to form test samples in said final holders; and

c. performing at least one of a plurality of assay methods on said test samples to generate test output data, wherein said automated workstation performs said assay methods.

2. The method of screening for microorganisms according to claim 1 wherein the step of providing a plurality of samples further comprises the steps of:

a. providing a sample source in a source holder; and

b. performing at least one of a plurality of inoculation methods to transfer a portion of said sample to said final holders, wherein said automated workstation performs said inoculation methods.

3. The method of screening for organisms according to claim 2 wherein said inoculation transfer methods, comprise the steps of distributing a portion of said sample from said sample source in said source holder to particular vessels of said pair of final holders in a manner in which creates specific patterns of distribution that said automated workstation maintains in memory and operates to effect said distribution under predetermined conditions.

4. The method of screening microorganisms according to claim 1 wherein the step of providing a chemical compound further comprises the steps of:

a. providing a chemical source of the chemical compound; and

b. performing at least one of a plurality of utility transfer methods to transfer said chemical compound to said pair of final holders, wherein said automated workstation performs said utility transfer methods.

5. The method of screening for organisms according to claim 4 wherein said utility transfer methods comprise the steps of distributing a portion of said chemical compound from said chemical source to said pair of final holders in a manner in which creates patterns of distribution that said automated workstation maintains in memory operates under predetermined conditions.

6. The method of screening for microorganisms according to claim 1 wherein said assay methods further comprise the step of detecting changes in the physical condition of said test samples and recording said detected changes to generate output data.

7. The method of screening for microorganisms according to claim 6 further comprising the step of calculating a growth curve based on said detected changes.

8. The method of screening for microorganisms according to claim 1 wherein said chemical compound is chromium.

9. The method of screening for microorganisms according to claim 1 wherein said automated workstation is a BioMek 1000.

10. The method of screening for microorganisms according to claim 1 wherein said plurality of samples is obtained from a contaminated site.

11. The method of screening for microorganisms according to claim 1 wherein said step of exposing said samples to said chemical compound to form test samples in said final holders is carried out so that said samples are exposed to said chemical compound in varying amounts.

12. A method of creating a growth curve while screening for microorganisms capable of detoxifying a compound, comprising the steps of:

a. i) an automated workstation;

ii) at least one chemical compound;

and iii) at least one sample;

b. measuring the optical density of at least one test sample;

c. calculating a growth curve for said microorganisms based on said optical density; and

d. performing operations on said test sample during said calculation step.

13. A method as recited in claim 12, prior to said measuring step, said method comprising the steps of:

a. providing;

i. a plurality of samples in a pair of final holders having separate vessels, each final holder separating the samples into said vessels, said samples suspected of containing microorganisms,

ii. a chemical compound, and

b. exposing said samples to said chemical compound to form said test samples in said final holders.

14. The method as recited in claim 13 wherein the step of providing a plurality of samples further comprises the steps of:

a. providing a sample source in a source holder; and

b. performing at least one of a plurality of inoculation methods to transfer a portion of said sample to said final holders, wherein said automated workstation performs said inoculation methods.

15. The method as recited in claim 14 wherein said inoculation transfer methods, comprise the steps of distributing a portion of said sample from said sample source in said source holder to particular vessels of said pair of final holders in a manner in which creates specific patterns of distribution that said automated workstation maintains in memory and operates to effect said distribution under predetermined conditions.

16. The method of screening microorganisms according to claim 12 wherein the step of providing a chemical compound farther comprises the steps of:

a. providing a chemical source of the chemical compound; and

b. performing at least one of a plurality of utility transfer methods to transfer said chemical compound to said pair of final holders.

17. The method as recited in claim 16 wherein said utility transfer methods comprise the steps of distributing a portion of said chemical compound from said chemical source to said pair of final holders in a manner in which creates patterns of distribution that said automated workstation maintains in said memory operates to effect under predetermined conditions.

18. The method as recited in claim 12 wherein said chemical compound is chromium.

20. The method as recited in claim 12 wherein said plurality of samples is obtained from a contaminated site.

19. The method as recited in 12 wherein said automated workstation is a BioMek 1000.

21. A method for screening microorganisms capable of detoxifying one or more compounds, comprising the steps of:

a) providing;

i) at least one sample suspected of containing said microorganisms; and

ii) at least one chemical compound; and

iii) an automated workstation;

b) exposing said sample to said chemical compound to produce a test sample; and

c) assaying said test sample for the presence of said chemical compound, wherein said assaying is performed by said automated workstation.

22. The method of claim 21, wherein said chemical compound is chromium.

23. The method of claim 21, wherein said sample is obtained from a contaminated site.

24. The method of claim 21, comprising the further step of providing optimal conditions for detoxification of said chemical compound by said microorganisms.

25. The method of claim 21, wherein said samples are exposed to varying concentrations of said chemical compound.

26. The method of claim 21, wherein said automated workstation is a BioMek 1000.

27. The method of claim 21, wherein said assay methods further comprise the step of detecting changes in the physical condition of said test samples and recording said detected changes to generate output data.

28. A method of creating a growth curve for microorganisms capable of detoxifying a compound, comprising the steps of:

a) providing;

i) a sample suspected of containing said microorganisms; and

ii) at least one chemical compound; and

iii) an automated workstation;

b) exposing said sample to said chemical compound to provide a test sample;

c) measuring the optical density of said test sample to provide an optical density determination, wherein said measuring is performed by said automated workstation; and

d) calculating a growth curve for said microorganisms from said optical density determination.

29. The method of claim 28, wherein said measuring said growth curve is calculated by said automated workstation.

30. The method of claim 28, wherein said automated workstation is a BioMek 1000.

31. The method of claim 28, wherein said chemical compound is chromium.

32. The method of claim 28, wherein said sample is obtained from a contaminated site.