Microbes and methods for remediation

A method for remediating a compound in a sample, the method comprising:

• providing killed cross-linked microbes comprising an exogenous polynucleotide comprising a coding region encoding a hydrolase that degrades a compound, wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed;
• and
• contacting a sample comprising the compound with the microbes under conditions effective to decrease the concentration of the compound in the sample relative to the concentration of the compound in a sample not contacted with the microbes.

A method for remediating a compound in a sample, the method comprising:

• providing killed cross-linked microbes comprising an exogenous polynucleotide comprising a coding region encoding a hydrolase that degrades an s-triazine, wherein the complement of the nucleotide sequence of the coding region hybridizes to the nucleotide sequence set forth at nucleotides 236 to 1660 of GenBank accession number U55933 in a solution containing 250 mM Na2HPO4, pH 7.4, 2 ml/liter 0.5 M EDTA, pH 8.0, and 10 grams/liter bovine serum albumin at 65° C. for at least 4 hours, followed by three washes for twenty minutes each at 65° C. in a solution containing 2×SSC and 0.1% SDS, and wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed;
• and
• contacting a sample comprising the s-triazine with the microbes under conditions effective to decrease the concentration of the s-triazine in the sample relative to the concentration of the s-triazine in a sample not contacted with the microbes.

A method for remediating a compound in a sample, the method comprising:

• providing killed cross-linked microbes comprising an exogenous polynucleotide comprising a coding region encoding a hydrolase that degrades atrazine, wherein the complement of the nucleotide sequence of the coding region hybridizes to the nucleotide sequence set forth at nucleotides 236 to 1660 of GenBank accession number U55933 in a solution containing 250 mM Na2HPO4, pH 7.4, 2 ml/liter 0.5 M EDTA, pH 8.0, and 10 grams/liter bovine serum albumin at 65° C. for at least 4 hours, followed by three washes for twenty minutes each at 65° C. in a solution containing 2×SSC and 0.1% SDS, and wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed;
• and
• contacting a sample comprising the atrazine with the microbes under conditions effective to decrease the concentration of the atrazine in the sample relative to the concentration of the atrazine in a sample not contacted with the microbes.

A method for remediating a compound in a sample, the method comprising:

• providing killed cross-linked microbes comprising an exogenous polynucleotide comprising a coding region encoding a hydrolase that degrades atrazine, wherein the nucleotide sequence of the coding region comprises nucleotides 236 to 1660 of GenBank accession number U55933, and wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed;
• and
• contacting a sample comprising the atrazine with the microbes under conditions effective to decrease the concentration of the atrazine in the sample relative to the concentration of the atrazine in a sample not contacted with the microbes.

A method for degrading an s-triazine in a sample, the method comprising:

• providing a cross-linked microbe comprising a polynucleotide comprising a coding region encoding a hydrolase that degrades an s-triazine;
• and
• contacting a sample comprising the s-triazine with the microbe under conditions effective to decrease the concentration of the s-triazine in the sample relative to the concentration of the s-triazine in a sample not contacted with the microbe.

A method for detoxifying a compound in a sample, the method comprising:

• providing killed cross-linked microbes comprising an exogenous polynucleotide comprising a coding region encoding a hydrolase that detoxifies a compound, wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed;
• and
• contacting a sample comprising the compound with the microbes under conditions effective to lower the toxicity of the compound in the sample relative to the toxicity of the compound in a sample not contacted with the microbes.

A method for detoxifying an s-triazine in a sample, the method comprising:

• providing killed cross-linked microbes comprising an exogenous polynucleotide comprising a coding region encoding a hydrolase that detoxifies an s-triazine, wherein the complement of the nucleotide sequence of the coding region hybridizes to the nucleotide sequence set forth at nucleotides 236 to 1660 of GenBank accession number U55933 in a solution containing 250 mM Na2HPO4, pH 7.4, 2 ml/liter 0.5 M EDTA, pH 8.0, and 10 grams/liter bovine serum albumin at 65° C. for at least 4 hours, followed by three washes for twenty minutes each at 65° C. in a solution containing 2×SSC and 0.1% SDS, and wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed;
• and
• contacting a sample comprising the s-triazine with the microbes under conditions effective to lower the toxicity of the s-triazine in the sample relative to the toxicity of the s-triazine in a sample not contacted with the microbes.

A cross-linked microbe comprising a polynucleotide comprising a coding region encoding a polypeptide that degrades an s-triazine, wherein the microbe has been killed.

Killed cross-linked microbes comprising a polynucleotide comprising a coding region encoding a hydrolase that degrades a compound, wherein the cross-linked microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed.

Killed cross-linked microbe comprising a polynucleotide comprising a coding region encoding a hydrolase that degrades an s-triazine, wherein the complement of the nucleotide sequence of the coding region hybridizes to the nucleotide sequence set forth at nucleotides 236 to 1660 of GenBank accession number U55933 in a solution containing 250 mM Na2HPO4, pH 7.4, 2 ml/liter 0.5 M EDTA, pH 8.0, and 10 grams/liter bovine serum albumin at 65° C. for at least 4 hours, followed by three washes for twenty minutes each at 65° C. in a solution containing 2×SSC and 0.1% SDS, and wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed.

Killed cross-linked microbes comprising a polynucleotide comprising a coding region encoding a hydrolase that degrades atrazine, wherein the complement of the nucleotide sequence of the coding region hybridizes to the nucleotide sequence set forth at nucleotides 236 to 1660 of GenBank accession number U55933 in a solution containing 250 mM Na2HPO4, pH 7.4, 2 ml/liter 0.5 M EDTA, pH 8.0, and 10 grams/liter bovine serum albumin at 65° C. for at least 4 hours, followed by three washes for twenty minutes each at 65° C. in a solution containing 2×SSC and 0.1% SDS, and wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed.

Killed cross-linked microbes comprising a polynucleotide comprising a coding region encoding a hydrolase that degrades atrazine, wherein the nucleotide sequence of the coding region comprises nucleotides 236 to 1660 of GenBank accession number U55933, and wherein the microbes retain at least about 30% hydrolase enzymatic activity compared to the hydrolase enzymatic activity of the microbes that are not killed.