Aspects of the invention include host cells that are engineered to produce benzylisoquinoline alkaloids (BIAs). The host cells include heterologous coding sequences for a variety of enzymes involved in synthetic pathways from starting compounds to BIAs of the host cell. Also provided are methods of producing the BIAs of interest by culturing the host cells under culture conditions that promote expression of enzymes encoded by the heterologous coding sequences of the host cells. Aspects of the invention further include compositions, e.g., host cells, starting compounds and kits, etc., that find use in methods of the invention.

A method for enzymatic preparation of fludarabine phosphate, comprising reaction of fludarabine with a high-energy phosphate compound under the action of deoxyribonucleic acid kinase. According to said method, acetate kinase and acetyl phosphate free acid or acetyl phosphate are also added. The technical problems present in the existing processes are successfully addressed by employing the enzymatic process to prepare the fludarabine phosphate. The usage of the high-energy phosphate compound is reduced by means of adding acetate kinase to recycle and regenerate a small amount of the high-energy phosphate compound, thereby reducing the generation of by-products having similar structures to the fludarabine phosphate, enhancing the operation convenience of purification steps in the industrial production of the fludarabine phosphate. The process is environment friendly, the reaction conditions are moderate, the cost is low, and the yield and the purity of the product obtained are high.

The present invention provides triazolone compounds of general formula (I):

##STR00001##

in which R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are as defined herein, methods of preparing said compounds, intermediate compounds useful for preparing said compounds, pharmaceutical compositions and combinations comprising said compounds and the use of said compounds for manufacturing pharmaceutical compositions for the treatment or prophylaxis of diseases, in particular of hyperproliferative disorders, as a sole agent or in combination with other active ingredients.

The disclosure provides biocatalysts that halogenate complex chemical compounds in specific and predictable ways. Also disclosed are halogenated complex organic compounds. The disclosure further provides methods for the halogenation of complex chemical compounds and methods of inhibiting the contraction of smooth muscle in mammals.

The present invention provides engineered ketoreductase and phosphite dehydrogenase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase and phosphite dehydrogenase enzymes, as well as polynucleotides encoding the engineered ketoreductase and phosphite dehydrogenase enzymes, host cells capable of expressing the engineered ketoreductase and phosphite dehydrogenase enzymes, and methods of using the engineered ketoreductase and phosphite dehydrogenase enzymes to synthesize a chiral catalyst used in the synthesis of antiviral compounds, such as nucleoside inhibitors. The present invention further provides methods of using the engineered enzymes to deracemize a chiral alcohol in a one-pot, multi-enzyme system.

A method of preparing neosaxitoxin in quantities of a purity sufficient to allow the compound to be used as an active pharmaceutical ingredient (API) is described. The method includes the reductive desulfonation of an unresolved mixture of gonyautoxin 1 (GTX1) and gonyautoxin 4 (GTX4).

Bacteria harbor an immense reservoir of potentially new and therapeutic small molecules in the form of “silent” biosynthetic gene clusters. These clusters can be identified bioinformatically but are at best sparingly expressed under normal laboratory growth conditions; their products are therefore not interrogated during bioactivity screening exercises. An estimated 80-90% of biosynthetic loci are silent, meaning that routine bioactivity screens miss the majority of microbial biosynthetic potential. Disclosed herein is a method that allows access to this vast hidden metabolome, thereby allowing researchers to screen the complete metabolomes of microorganisms in the search of new therapeutic leads. The disclosed approach, “Bioactivity-HiTES”, broadly activates the secondary metabolomes of bacteria and links the cryptic metabolites produced to a desired biological activity. Using the disclosed method, induction of cryptic antibiotics was detected in all four actinomycete bacterial strains that were tested as proof-of-concept. Follow-up in two cases demonstrated the production of two new antibiotics: In one case, the taylorflavins, pyrimidine antibiotics that harbor selective growth-inhibitory activity toward Gram-negative bacteria, were identified. For example, taylorflavin B shows potent minimal inhibitory concentration toward Neisseria gonorrhoeae and Acinetobacter baumanii, but not against a panel of Gram-positive bacteria. In the second case, the lanthipeptide cebulantin, which is specific toward Gram-negative Vibrio pathogens, but does not affect the growth of Gram-positive bacteria tested, was identified. These compounds, taylorflavins and cebulantin, may serve as useful leads in the future. At the same time, Bioactivity-HiTES may be applied broadly to identify cryptic metabolites with the desired biological properties.

The present invention relates to processes to make neosaxitoxin, and analogues and variants thereof, and intermediates in the production of neosaxitoxin in recombinant host cells. Neosaxitoxin and the analogues and variants thereof may be used in the production of pharmaceutical compositions.

The present invention discloses processes for obtaining a color from an algal biomass. The process includes subjecting an algal biomass to cavitation, thus at least partially disrupting cells of the algal biomass and extracting color from the disrupted, algal biomass.

Provided herein are microorganisms that include one or more heterologous nucleic acid selected from the group of a sequence encoding a 7-methylxanthosine synthase, a sequence encoding a theobromine synthase; and a sequence encoding a caffeine synthase, where the microorganism is capable of producing one or more purine alkaloid in a culture medium, when the microorganism is cultured under conditions sufficient to produce the one or more purine alkaloid. Also provided compositions and kits that include at least one of these microorganisms, and methods of producing one or more purine alkaloid that include culturing one of these microorganisms under conditions sufficient to produce the one or more purine alkaloid.