Single-step affinity and cost-effective purification of recombinant proteins using the Sepharose-binding lectin-tag from the mushroom Laetiporus sulphureus as fusion partner.

Previous research showed that a lectin from the mushroom Laetiporus sulphureus, designed LSL, bound to Sepharose and could be eluted by lactose. In this study, by taking advantage of the strong affinity of LSL-tag for Sepharose, we developed a single-step purification method for LSL-tagged fusion proteins. We utilized unmodified Sepharose-4B as a specific adsorbent and 0.2 M lactose solution as an elution buffer. Fusion proteins of LSL-tag and porcine circovirus capsid protein, designated LSL-Cap was recovered with purity of 90 ± 4%, and yield of 87 ± 3% from crude extract of recombinant Escherichia coli. To enable the remove of LSL-tag, tobacco etch virus (TEV) protease recognition sequence was placed downstream of LSL-tag in the expression vector, and LSL-tagged TEV protease, designated LSL-TEV, was also expressed in E. coli., and was recovered with purity of 82 ± 5%, and yield of 85 ± 2% from crude extract of recombinant E. coli. After digestion of LSL-tagged recombinant proteins with LSL-TEV, the LSL tag and LSL-TEV can be easily removed by passing the digested products through the Sepharose column. It is of worthy noting that the Sepharose can be reused after washing with PBS. The LSL affinity purification method enables rapid and inexpensive purification of LSL-tagged fusion proteins and scale-up production of native proteins.

Heavy metal pollution in coastal areas of South China: a review.

Coastal areas of South China face great challenges due to heavy metal contamination caused by rapid urbanization and industrialization. In this paper, more than 90 articles on levels, distributions, and sources of heavy metals in sediments and organisms were collected to review the status of heavy metal pollution along coastal regions of South China. The results show that heavy metal levels were closely associated with local economic development. Hong Kong and the Pearl River Estuary were severely contaminated by heavy metals. However, concentrations of heavy metals in sediments from Hong Kong have continually decreased since the early 1990 s. High levels of heavy metals were found in biota from Lingdingyang in Guangdong province. Mollusks had higher concentrations of heavy metals than other species. Human health risk assessments suggested that levels of heavy metals in some seafood from coastal areas of South China exceeded the safety limit.