A microorganism including a foreign gene encoding a protein having a hydroxylase activity that reduces the concentration of CH.sub.nF.sub.4-n (n is an integer of 0 to 3) in a sample, as well as a composition including the microorganism or lysate thereof, and a method of reducing the concentration of CH.sub.nF.sub.4-n in a sample using the microorganism or lysate.

A first aspect of the invention relates to a non-covalently-HaloTag-binding compound characterized by the general formula D-L-T (I), wherein D is or comprises a functional moiety Z, particularly a fluorescent dye, or D is a linkable moiety (i.e. a moiety that can be coupled to other functional groups), L is a linear linker of 10-15 atoms in length, and T is a moiety selected from the group comprising methylamine, methylsulfonamide, acetamide, or their respective fluorinated analogues, azide, or hydroxyl. Another aspect of the invention relates to a HaloTag variant wherein position D106 of the HaloTag7 sequence is exchanged for a proteinogenic amino acid different from D. The variant has a different binding specificity for HaloTag substrates compared to the non-variant Halotag polypeptide. Yet another aspect of the invention relates to kits comprising polypeptides or nucleic acids and the non-covalently-HaloTag-binding compound according to the invention.

The present invention relates to the fields of life sciences, micro-organisms and degradation of polyolefin polymers. Specifically, the invention relates to an isolated specific enzyme or a fragment thereof, wherein said enzyme or fragment is capable of degrading a polyolefin, and to a micro-organism or a host cell comprising the enzyme or a fragment thereof. Also, the present invention relates to a polynucleotide encoding the enzyme or fragment thereof, and to an expression vector or plasmid comprising the polynucleotide of the present invention. And still, the present invention relates to use of the enzyme, fragment, micro-organism, host cell, polynucleotide, expression vector or plasmid of the present invention for degrading a polyolefin: to a method of degrading a polyolefin with the specific enzyme or a fragment thereof: and to a method of producing the enzyme or fragment thereof of the present invention.

A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

Provided are a dehalogenase variant, a polynucleotide encoding the dehalogenase variant, a recombinant microorganism including a genetic modification that increases dehalogenase activity, a composition including the recombinant microorganism, and a method of reducing a concentration of fluorinated methane using the recombinant microorganism.

Provided are a microorganism having activity in reducing a concentration of a fluorine-containing compound in a sample, a recombinant microorganism including a gene derived from the microorganism, and a method of reducing the concentration of the fluorine-containing compound in the sample by using the microorganism or recombinant microorganism.

Provided is a microorganism including a gene encoding a protein having a dehalogenase activity, a composition for using in reducing a concentration of fluorinated methane in a sample, the composition including the microorganism including the gene encoding the protein having the dehalogenase activity, and a method of reducing the concentration of fluorinated methane in the sample.

A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

Provided are a recombinant microorganism including a first genetic modification that increases activity of haloalkane dehalogenase (HAD) and a second genetic modification that increases expression of at least one of RcIR, RcIA, RcIB, and RcIC; a composition for reducing a concentration of fluorinated methane in a sample, wherein the composition includes the recombinant microorganism; and a method of reducing a concentration of fluorinated methane in a sample.