The present disclosure describes the engineering of microbial cells for fermentative production of histamine and provides novel engineered microbial cells and cultures, as well as related histamine production methods.

Described herein are variants of acetylserotonin O-methyltransferase (ASMT) as well as vectors and recombinant microbial host cells expressing such ASMT variant and their use in producing melatonin and related compounds. Preferred ASMT variants provide for a higher turnover of N-acetylserotonin into melatonin.

Described herein are variants of acetylserotonin O-methyltransferase (ASMT) as well as vectors and recombinant microbial host cells expressing such ASMT variant and their use in producing melatonin and related compounds. Preferred ASMT variants provide for a higher turnover of N-acetylserotonin into melatonin.

Integral membrane proteins capable of transporting melatonin and other indole-derivatives across biological membranes, and uses thereof.

Integral membrane proteins capable of transporting melatonin and other indole-derivatives across biological membranes, and uses thereof.

A microorganism of the present invention is Dirkmeia churashimaensis (NITE BP-03054), Papiliotrema flavescens (NITE BP-03051), Papiliotrema flavescens (NITE BP-03052), or Apiotrichum porosum (NITE BP-03053).

A method of culturing human induced pluripotent stem cells includes inoculating human induced pluripotent stem cells in a culture medium at an inoculation density of 1.0×10.sup.4 to 1.0×10.sup.6 cells/cm.sup.2 in a culture vessel and subjecting the human induced pluripotent stem cells to two-dimensional culturing. A method of producing cerebral organoids includes culturing a culture of the human induced pluripotent stem cells obtained by the method of culturing human induced pluripotent stem cells in a culture medium containing a BMP inhibitor and a transforming growth factor β (TGFβ) inhibitor to form cell aggregates, culturing the cell aggregates in a culture medium containing a Wnt signal transduction pathway potentiator and an extracellular matrix, and subjecting the culturing obtained in the culturing the cell aggregates to spinner culturing.

The present invention provides a polypeptide tag and its application in the synthesis of pharmaceutical chemicals, the recombinant nitrilase was obtained by connecting a polypeptide tag to the N-terminus of the amino acid sequence of the nitrilase; wherein amino acids at both ends of the polypeptide tag are uncharged glycine G, and the rest are a random combination of any one or more of glycine G, histidine H, glutamic acid E, aspartic acid D, lysine K and arginine R; The activity of the recombinant nitrilase in the preparation of 1-cyanocyclohexyl acetic acid is up to 3034.7 U/g dcw, the polypeptide tag significantly improves the soluble expression of nitrilase, and the whole cell catalyst hydrolyzes 1M substrate with the same concentration 30 minutes faster than the mother enzyme. The method provided by the present invention can also be used for the biocatalytic reaction of other pharmaceutical intermediates as the substrate catalyzed by the nitrilase, improving the activity of the whole cell catalyst in reaction, and also improving the solubility of other types of nitrilases and the activity of the corresponding whole cell catalysts.

Provided are non-naturally occurring nucleic acids comprising a sequence encoding an enzyme or regulatory protein in tryptamine metabolism. Also provided are a recombinant microorganisms expressing the enzyme or regulatory protein. Methods of expressing the enzyme or regulatory protein are additionally provided.

The present invention is directed to a process for preparing a compound of formula I-11 through multiple-step reactions: ##STR00001##