In September 2002, the American Association of Colleges of Pharmacy (AACP) Council of Deans and Pharmacy Practice Section appointed this Task Force and charged it to develop a white paper addressing the role of colleges and schools in developing and administering residency programs. Specifically, the issues to be addressed included:1. What role should colleges and schools of pharmacy play in developing new residencies? Is there a national need at this time for new ones? If so, what should collegiate responsibility be? What are potential models of investment to help develop new residencies?Corollary: What role does residency training play in preparing future faculty? Are there educational experiences that should be incorporated into residency programs that prepare future faculty?2. Explore the postgraduate medical training model that involves schools holding the accreditation for a program and schools then approving the specific training sites. Such a model is already possible for community pharmacy residencies. Should it be applied to other types of pharmacy residency training? What are the advantages and disadvantages of such a model?3. What needs to be done to make available maximum resources for residency training?4. How can colleges partner with residency sites in residency research projects and encourage some residents to seek additional training in order to assume faculty positions? BACKGROUNDPharmacy residency programs are desirable for advanced patient care practice and future pharmacy leaders. Such programs are estimated to be equivalent to three to five years of work experience to prepare pharmacists for a variety of practice settings. Individuals with residency training can assist colleges and schools of pharmacy in several ways: (a) as full time faculty; (b) as preceptors for the experiential portion of the program; (c) as administrators at hospitals and clinics who can partner with colleges of pharmacy on a common educational mission; (d) as professional leaders on political advocacy issues that can enhance resources to support entry level and advanced pharmacy education; and (e) as pharmacist role models of patient care practice. For this reason, it is essential that colleges of pharmacy assume a significant role in residency training.Colleges and schools of pharmacy are also facing faculty shortages in departments/divisions of pharmacy practice.1 AACP contacted 84 member institutions to complete a survey in December 2002. Responses were received from 67 (79%) colleges/schools, which reported a total of 417 vacant teaching positions, or an average of 6 vacancies per college/school. Of the vacancies, 53.4% were in pharmacy practice and 45.6% were in pharmaceutical sciences. Of the vacancies, 94% were full time faculty positions. 2 The shortages are thought to be due to an inadequate supply of potential faculty, problems with faculty retention, and increased demand for faculty. Multiple etiologies have been proposed including lower salaries of faculty positions compared with industry, hospital, or c...

A crotonyl-CoA reductase (EC 1.3.1.38, acyl-CoA :NADP' trans-2-oxidoreductase) catalyzing the conversion of crotonyl-CoA to butyryl-CoA has been purified and characterized from Streptomyces collinus. This enzyme, a dimer with subunits of identical mass (48 kDa), exhibits a K,, = 18 pM for crotonylCoA and 15 pM for NADPH. The enzyme was unable to catalyze the reduction of any other enoyl-CoA thioesters or to utilize NADH as an electron donor. A highly effective inhibition by straight-chain fatty acids ($ = 9.5 pM for palmitoyl-CoA) compared with branched-chain fatty acids (K,>400 pM for isopalmitoyl-CoA) was observed. All of these properties are consistent with a proposed role of the enzyme in providing butyryl-CoA as a starter unit for straight-chain fatty acid biosynthesis. The crotonyl-CoA reductase gene was cloned in Escherichia coli. This gene, with a proposed designation of ccr, is encoded in a 1344-bp open reading frame which predicts a primary translation product of 448 amino acids with a calculated molecular mass of 49.4 kDa. Several dispersed regions of highly significant sequence similarity were noted between the deduced amino acid sequence and various alcohol dehydrogenases and fatty acid synthases, including one region that contains a putative NADPH binding site. The ccr gene product was expressed in E. coli and the induced crotonyl-CoA reductase was purified tenfold and shown to have similar steady-state kinetics and electrophoretic mobility on sodium dodecyl sulfate/polyacrylamide to the native protein.

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We report the cloning of the gene encoding the 1-cyclohexenylcarbonyl coenzyme A reductase (ChcA) of Streptomyces collinus, an enzyme putatively involved in the final reduction step in the formation of the cyclohexyl moiety of ansatrienin from shikimic acid. The cloned gene, with a proposed designation of chcA, encodes an 843-bp open reading frame which predicts a primary translation product of 280 amino acids and a calculated molecular mass of 29.7 kDa. Highly significant sequence similiarity extending along almost the entire length of the protein was observed with members of the short-chain alcohol dehydrogenase superfamily. The S. collinus chcA gene was overexpressed in Escherichia coli by using a bacteriophage T7 transient expression system, and a protein with a specific ChcA activity was detected. The E. coli-produced ChcA protein was purified and shown to have similar steady-state kinetics and electrophoretic mobility on sodium dodecyl sulfate-polyacrylamide gels as the enoyl-coenzyme A reductase protein prepared from S. collinus. The enzyme demonstrated the ability to catalyze, in vitro, three of the reductive steps involved in the formation of cyclohexanecarboxylic acid. An S. collinus chcA mutant, constructed by deletion of a genomic region comprising the 5 end of chcA, lost the ChcA activity and the ability to synthesize either cyclohexanecarboxylic acid or ansatrienin. These results suggest that chcA encodes the ChcA that is involved in catalyzing multiple reductive steps in the pathway that provides the cyclohexanecarboxylic acid from shikimic acid.

A number of microorganisms were selected from soil and sediment samples which were known to have been previously exposed to nitrate ester contaminants. The two most effective bacteria for transforming glycerol trinitrate (GTN) were identified as Bacillus thuringiensis/cereus and Enterobacter agglomerans. For both isolates, denitration activities were expressed constitutively and GTN was not required for induction. Dialysis of cell extracts from both isolates did not affect denitration, which indicates that dissociable and depletable cofactors are not required for denitration. With thin-layer chromatography and high-performance liquid chromatography, the denitration pathway for both isolates was shown to be a sequential denitration of GTN to glycerol dinitrate isomers, glycerol mononitrate isomers, and ultimately to glycerol. GTN was observed to be completely converted to glycerol during a long-term incubation of cell extracts.

A novel NADPH-dependent enoyl reductase, catalyzing the conversion of 1-cyclohexenylcarbonyl coenzyme A (1-cyclohexenylcarbonyl-CoA) to cyclohexylcarbonyl-CoA, was purified to homogeneity from Streptomyces coUinus. This enzyme, a dimer with subunits of identical Mr (36,000), exhibits a Km of 1.5 0.3 ,uM for NADPH and 25 + 3 ,uM for 1-cyclohexenylcarbonyl-CoA. It has a pH optimum of 7.5, is most active at 30°C, and is inhibited by both divalent cations and thiol reagents. Two internal peptide sequences were obtained.

Currently, there has been limited use of genetic engineering for waste treatment. In this work, we are developing a procedure for the in situ treatment of toxic organophosphate wastes using the enzyme parathion hydrolase. Since this strategy is based on the use of an enzyme and not viable microorganisms, recombinant DNA technology could be used without the problems associated with releasing genetically altered microorganisms into the environment. The gene coding for parathion hydrolase was cloned into a Streptomyces lividans, and this transformed bacterium was observed to express and excrete this enzyme. Subsequently, fermentation conditions were developed to enhance enzyme production, and this fermentation was scaled-up to the pilot scale. The cell-free culture fluid (i.e., a nonpurified enzyme solution) was observed to be capable of effectively hydrolyzing organophosphate compounds under laboratory and simulated in situ conditions.