Adsorption and binding of the transgenic plant proteins, human serum albumin, β-glucuronidase, and Cry3Bb1, on montmorillonite and kaolinite: Microbial utilization and enzymatic activity of free and clay-bound proteins

Abstract Human serum albumin (HSA), β-glucuronidase (GUS), and the Cry3Bb1 protein from Bacillus thuringiensis subsp. kumamotoensis are expressed by genetically-modified plants. Commercial samples of these proteins adsorbed and bound rapidly on the clay minerals, kaolinite (K) and montmorillonite (M). Adsorption increased as the concentration of protein increased and then reached a plateau. The greatest amount of adsorption and binding occurred with the Cry3Bb1 protein, of which there was no desorption: 6.7 ±0.21 μg adsorbed and bound μg− 1 of M; 2.1 ± 0.39 μg adsorbed and bound μg− 1 of K. With GUS, 2.2 ± 0.29 μg adsorbed and 1.7 ±0.21 μg bound μg− 1 of M; 1.5 ± 0.28 μg adsorbed and 1.0 ± 0.03 μg bound μg− 1 of K. HSA was adsorbed and bound the least: 1.2 ±0.04 μg adsorbed and 0.8 ± 0.05 μg bound μg− 1 of M; 0.4 ± 0.05 μg adsorbed and 0.4 ± 0.03 μg bound μg− 1 of K. However, X-ray diffraction analyses indicated that only HSA intercalated M, and none of the proteins intercalated K, a nonswelling clay. When bound, the proteins were not utilized for growth by mixed cultures of soil microorganisms, whereas the cultures readily utilized the free (i.e., not adsorbed or bound) proteins as sources of carbon and energy. The enzymatic activity of GUS was significantly enhanced when bound on the clay minerals. These results indicated that recombinant proteins expressed by transgenic plants could persist and function in soil after release in root exudates and from decaying plant residues as the result of the protection provided against biodegradation by binding on clay minerals.