Bioencapsulation of living bacteria (Escherichia coli) with poly(silicate) after transformation with silicatein-alpha gene.
Bioencapsulation is an intriguing way to immobilize biological materials, including cells, in silica, metal-oxides or hybrid sol-gel polymers. Until now only the sol-gel precursor technology was utilized to immobilize bacteria or yeast cells in silica. With the discovery of silicatein, an enzyme from demosponges that catalyzes the formation of poly(silicate), it became possible to synthesize poly(silicate) under physiological (ambient) conditions. Here we show that Escherichia coli can be transformed with the silicatein gene, its expression level in the presence of isopropyl beta-D-thiogalactopyranoside (IPTG) can be efficiently intensified by co-incubation with silicic acid. This effect could be demonstrated on the level of recombinant protein synthesis as well as by immunostaining analysis. The heterologously produced silicatein is enzymatically active, as confirmed by staining with Rhodamine 123 (formation for poly[silicate] from silicic acid) and by reacting free silicic acid with the beta-silicomolybdato color system. Electron microscopic analysis revealed that the bacteria that express silicatein form a viscous cover around them when growing in the presence of silicic acid. Finally, we demonstrate that the growth kinetics of E. coli remains unaffected whether or not the bacteria had been transformed with silicatein or grown in medium, supplemented with silicic acid. It is concluded that silicatein-mediated encapsulation of bacteria with silica might improve, extend and optimize the range of application of bacteria for the production of recombinant protein.
Müller
Werner E G
WE
Institut für Physiologische Chemie, Abteilung Angewandte Molekularbiologie, Universität, Duesbergweg 6, D-55099 Mainz, Germany. wmuller@uni-mainz.de
Engel
Sylvia
S
Wang
Xiaohong
X
Wolf
Stephan E
SE
Tremel
Wolfgang
W
Thakur
Narsinh L
NL
Krasko
Anatoli
A
Divekar
Mugdha
M
Schröder
Heinz C
HC
D016667:Bacterial Capsules / Q000737:chemistry*; D002403:Cathepsins / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D050296:Microbial Viability; D008855:Microscopy, Electron, Scanning; D017640:Silicates / Q000737:chemistry*; D012964:Sodium / Q000737:chemistry; D019076:Transgenes / Q000235:genetics*
D017640:Silicates; C112571:silicatein alpha; D012964:Sodium; D002403:Cathepsins