Enzymatic deprotection of amines and hydroxides

Details Enzymatic deprotection of amines and hydroxides Enzymatic deprotection of amines and hydroxides

2001-12-14

2004-12-07

Weber, Jon P (Department: 1651)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

C435S106000, C435S117000

Reexamination Certificate

active

06828119

ABSTRACT:

The present invention relates to mild, enzyme driven methods for removing amine and hydroxide protecting groups.
N-carbobenzyloxy (N-CBZ) group is commonly used to protect amino and hydroxide groups during organic synthesis. Other similar “carbamate” protecting groups are also used to protect amino groups. Chemical deprotection is usually achieved by methods such as hydrogenation with Palladium catalyst. However, if other groups are present that are susceptible to the deprotection condition (for example, sulfur during hydrogenation), alternative methods of deprotection are necessary. It has now been discovered that microorganisms can be readily isolated from soil samples using a selection technique of the invention to produce an enzyme activity effective to specifically release such protecting groups. Thus, an enzymatic method of deprotection conducted under mild conditions (e.g., aqueous medium at room temperature and atmospheric pressure) can be used which avoids damaging any susceptible or potentially susceptible groups.
SUMMARY OF THE INVENTION
The invention provides a method of deprotecting a hydroxide or amine protected with a group of formula
ArC*(R)H—(CH
2
)
n
—O—C(═O)—
where the substituents are as described below, the method comprising: contacting the protected hydroxide or amine with an enzyme effective to remove the protecting group; and recovering the amine. Also provided is a method of isolating a bacteria producing an enzyme effective to remove a protecting group comprising: growing prospective bacteria on a medium having a growth selective amount of an amine compound that is protected as above; and isolating bacteria that grow on said medium.
The invention further provides a method of resolving a desired enantiomer of an amine or hydroxide linked to a chiral carbon. The amine or hydroxide protected with such a group is stereo-specifically hydrolyzed with the method of the invention. The desired enantiomer is either that hydrolyzed or that resistant to hydrolysis.
In one embodiment, the contacting step effectuates the following reaction:
where Pr— is the above-described protecting group. In another embodiment, the contacting effectuates the following reaction:
In yet another embodiment, the contacting effectuates the following reaction:


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