Tsinghua University

California Institute of Technology

The South China University of Technology

Zhanglin Lin

Qing Zhao

Lei Xing

Bihong Zhou

Frances H Arnold

Xu Wang

Wei Wu

Wanghui Xu

Lingli Zeng

Tingting Wang

Hyun Joo

The present disclosure describes a Mtu &#x394;I-CM intein variant containing one or more mutations or a biologically active fragment thereof, and a method for producing and purifying a molecule of interest using the intein variant. Further described are isolated fusion proteins comprising the intein variant and a tag and a molecule of interest. Also described are expression systems for expressing the intein variant as well as polypeptide screening methods employing the intein variant.

Fusion proteins comprising a Mtu &#x394;I-CM intein variant and methods of protein purification

The present invention relates to a method for production and purification of polypeptides. In particular, the present invention relates to a fusion protein comprising a solubility-enhancing peptide tag moiety, a self-aggregating peptide moiety and a moiety of target peptide and to a method for production and purification of target peptides through expressing said fusion protein.

Methods for production and purification of polypeptides

A method for detecting the presence of an oxygenated compound which is produced when a substrate is reacted with an oxygenase for the substrate. The method involves reacting a coupling enzyme with the oxygenated compound to form a polymeric oxygenated compound which is fluorescent or luminescent. Measurement of the fluorescence or luminescence of the polymeric oxygenated compound provides indirect detection of the oxygenated compound produced by reaction of the oxygenase with the substrate. The method is carried out in a whole cell environment wherein the cell is transformed to express both the oxygen a set being screened and the coupling enzyme. The method can be used to measure the activity of monooxygenases and dioxygenases on aromatic substrates. The method is amenable to large scale screening of enzyme mutants to isolate those with maximum oxygenase activity.

Oxygenase enzymes and screening method

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Beijing, China

Zhanglin Lin