Drawing inspiration from nature to develop anti-fouling coatings: the development of biomimetic polymer surfaces and their effect on bacterial fouling

The development of self-cleaning biomimetic surfaces has the potential to be of great benefit to human health, in addition to reducing the economic burden on industries worldwide. Consequently, this study developed a biomimetic wax surface using a moulding technique which emulated the topography of the self-cleaning Gladiolus hybridus (Gladioli) leaf. A comparison of topographies was performed for unmodified wax surfaces (control), biomimetic wax surfaces, and Gladioli leaves using optical profilometry and scanning electron microscopy. The results demonstrated that the biomimetic wax surface and Gladioli leaf had extremely similar surface roughness parameters, but the water contact angle of the Gladioli leaf was significantly higher than the replicated biomimetic surface. The self-cleaning properties of the biomimetic and control surfaces were compared by measuring their propensity to repel Escherichia coli and Listeria monocytogenes attachment, adhesion, and retention in mono-and co-culture conditions. When the bacterial assays were carried out in monoculture, the biomimetic surfaces retained fewer bacteria than the control surfaces. However, when using co-cultures of the bacterial species, only following the retention assays were the bacterial numbers reduced on the biomimetic surfaces. The results demonstrate that such surfaces may be effective in reducing biofouling if used in the appropriate medical, marine, and industrial scenarios. This study provides valuable insight into the anti-fouling physical and chemical control mechanisms found in plants, which are particularly appealing for engineering purposes

Export of functional Streptomyces coelicolor alditol oxidase to the periplasm or cell surface of Escherichia coli and its application in whole-cell biocatalysis

Streptomyces coelicolor A3(2) alditol oxidase (AldO) is a soluble monomeric flavoprotein in which the flavin cofactor is covalently linked to the polypeptide chain. AldO displays high reactivity towards different polyols such as xylitol and sorbitol. These characteristics make AldO industrially relevant, but full biotechnological exploitation of this enzyme is at present restricted by laborious and costly purification steps. To eliminate the need for enzyme purification, this study describes a whole-cell AldO biocatalyst system. To this end, we have directed AldO to the periplasm or cell surface of Escherichia coli. For periplasmic export, AldO was fused to endogenous E. coli signal sequences known to direct their passenger proteins into the SecB, signal recognition particle (SRP), or Twin-arginine translocation (Tat) pathway. In addition, AldO was fused to an ice nucleation protein (INP)-based anchoring motif for surface display. The results show that Tat-exported AldO and INP-surface-displayed AldO are active. The Tat-based system was successfully employed in converting xylitol by whole cells, whereas the use of the INP-based system was most likely restricted by lipopolysaccharide LPS in wild-type cells. It is anticipated that these whole-cell systems will be a valuable tool for further biological and industrial exploitation of AldO and other cofactor-containing enzymes.

Functional Expression of Spider Neurotoxic Peptide Huwentoxin-I in E. coli

The coding sequence of huwentoxin-I, a neurotoxic peptide isolated from the venom of the Chinese spider Ornithoctonus huwena, was amplified by PCR using the cDNA library constructed from the spider venom glands. The cloned fragment was inserted into the expression vector pET-40b and transformed into the E. coli strain BL21 (DE3). The expression of a soluble fusion protein, disulfide interchange protein (DsbC)-huwentoxin-I, was auto-induced in the periplasm of E. coli in the absence of IPTG. After partial purification using a Ni-NTA column, the expressed fusion protein was digested using enterokinase to release heteroexpressed huwentoxin-I and was further purified using RP-HPLC. The resulting peptide was subjected to gel electrophoresis and mass spectrometry analysis. The molecular weight of the heteroexpressed huwentoxin-I was 3750.69, which is identical to that of the natural form of the peptide isolated from spider venom. The physiological properties of the heteroexpressed huwentoxin-I were further analyzed using a whole-cell patch clamp assay. The heteroexpressed huwentoxin-I was able to block currents generated by human Nav1.7 at an IC50 of 640 nmole/L, similar to that of the natural huwentoxin-I, which is 630 nmole/L