This disclosure describes recombinant Megasphaera microbes designed to include increased consumption of acetate, increased carbon flux to butyryl-CoA and/or hexanoyl-CoA, increased production of butyrate and/or hexanoate, or a combination thereof, than a comparable control. This disclosure also describes methods that generally include growing such recombinant microbes under conditions effective for the recombinant microbes to consume greater amounts of acetate, produce increased amounts of butyryl-CoA and/or hexanoyl-CoA, produce increased amounts of butyrate and/or hexanoate, or a combination thereof.

The present invention relates to the genetic modification of bacteria of the genus Clostridium, typically solventogenic bacteria of the genus Clostridium, in particular bacteria possessing in the wild type a gene encoding an amphenicol-O-acetyltransferase. It thus relates to methods, tools and kits allowing such a genetic modification, in particular the removal or modification of a sequence encoding or controlling the transcription of an amphenicol-O-acetyltransferase, to the genetically modified bacteria obtained and to uses thereof, in particular for producing a solvent, preferably on an industrial scale.

A modified chloramphenicol acetyltransferase comprising a tyrosine residue 20 having a phenylalanine (Y20F) mutation, a microorganism harboring the modified chloramphenicol acetyltransferase, and a method of producing ester by feeding the microorganism are disclosed. The method includes providing the microorganism harboring a modified chloramphenicol acetyltransferase in an environment suitable for the microorganism to produce an ester and feeding the microorganism (i) a sugar or a cellulose, and (ii) an alcohol and/or a carboxylic acid.

A method is provided for increasing an immunological response to a target antigen in an animal by administering an immunogenic amount of a vaccine comprising a polypeptide conjugate comprising the target antigen conjugated to a carrier polypeptide by means of a linker polypeptide which is rich in predicted linear B-cell epitopes.

A method for biologically producing acetin such as monoacetin, diacetin, or triacetin according to an embodiment of the present disclosure includes reacting acetyl-CoA with glycerol in the presence of a first O-acetyl transferase to obtain the acetin. With the method, acetin which is sustainable and safe, and has more excellent quality while not causing environmental pollution may be obtained.

The design, assembly, and use of novel sequences comprising targeting and insertion sites for site-specific bacterial transposons are disclosed. One aspect relates to a nucleotide sequence comprising an attachment site for a site-specific transposon operably-linked to a screenable or selectable marker sequence, wherein said marker sequence encodes one or more active or inactive polypeptides capable of conferring a screenable or selectable phenotype upon a cell comprising the marker sequence, wherein insertion of the site-specific transposon into the attachment site changes the phenotype of a cell comprising the screenable or selectable marker sequence. High and low copy number vectors comprising the sequences, designated synthemids, including plasmids capable of propagating in bacteria, and shuttle vectors, capable of propagating in bacteria and a eukaryotic host cell or two types of bacteria by means of distinct replicons, are also disclosed. Related aspects include the design and assembly of synthetic insect and mammalian virus shuttle vectors, including shuttle vectors comprising segments of a double-stranded DNA virus, such as a baculovirus, which propagates in insect cells, or a herpesvirus, an adenovirus, or a pox virus, which propagate in mammalian cells. Other aspects relate to use of modified vectors to express polypeptides for use as therapeutic drug products, as vaccines, or as components of cell or gene therapy vector systems, and in model and crop plant cells, tissues, and whole plants to facilitate the basic and applied studies leading to improved food products, and as tools advancing the interests of institutions involved in industrial and environmental biotechnology.

Disclosed herein are enzymatic and modified host cell methods for prepared modified indole alkaloids. Also disclosed herein are modified indole alkaloids and their therapeutic use for treating diseases and disorders.

A method is provided for increasing an immunological response to a target antigen in an animal by administering an immunogenic amount of a vaccine comprising a polypeptide conjugate comprising the target antigen conjugated to a carrier polypeptide by means of a linker polypeptide which is rich in predicted linear B-cell epitopes.

A fusion polypeptide, comprising an inclusion body tag fused to at least one polypeptide of interest, is provided. The inclusion body tag comprises an inclusion body forming (IBF) amino acid sequence selected from X.sub.14X.sub.15AX.sub.17X.sub.18X.sub.19GLTVA GMLG and sequences having at least 71% identity thereto. Also provided are inclusion bodies comprising the fusion polypeptide, as well as nucleic acids, expression vectors, host cells and methods for its production.

A method is provided for increasing an immunological response to a target antigen in an animal by administering an immunogenic amount of a vaccine comprising a polypeptide conjugate comprising the target antigen conjugated to a carrier polypeptide by means of a linker polypeptide which is rich in predicted linear B-cell epitopes.