Surface display of lipolytic enzyme, Lipase A and Lipase B of Bacillus subtilis on the Bacillus subtilis spore
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
Authors: Kim, Junehyung
Abstract
For the enhancement of lipase stability in organic solvent containing reaction, live immobilization method, using Bacillus subtilis spore as a display vehicle was attempted. Bacillus subtilis coat protein cotE was used as an anchoring motif for the display of lipA and lipB of Bacillus subtilis. Using this motif, lipolytic enzyme Lipase A and Lipase B were functionally displayed on the surface of Bacillus subtilis spore. Purified spore displaying CotE-LipB fusion protein showed higher lipolytic activity compared to that of CotE-LipA fusion protein. The surface localization of Lipase B was verified with flow cytometry and protease accessibility experiment. Spore displayed lipase retained its activity against acetone and benzene which completely deactivated free soluble lipase in the same reaction condition.
Oral HPV prevalence assessment by Linear Array vs. SPF10 PCR-DEIA-LiPA(25) system in the HPV Infection in Men (HIM) study
PAPILLOMAVIRUS RESEARCH
Authors: Bettampadi, Deepti; Sirak, Bradley A.; Fulp, William J.; Abrahamsen, Martha; Villa, Luisa L.; Lazcano-Ponce, Eduardo; Salmeron, Jorge; Isaacs-Soriano, Kimberly A.; Baggio, Maria L.; Quiterio Trenado, Manuel; Giuliano, Anna R.
Abstract
Introduction: Oral human papillomavirus (HPV) attributable oropharyngeal cancers are on the rise in many countries. Oral HPV infections among healthy individuals are commonly detected using oral gargle samples. However, the optimal method for HPV genotyping oral gargle specimens in research studies has not been previously evaluated. Materials and methods: Oral gargle samples from 1455 HPV Infection in Men (HIM) study participants were HPV genotyped using two different methods: Linear Array and the SPF10 PCR-DEIA-LiPA(25). The sensitivity of the two tests for detecting individual HPV types and grouped HPV types, high-risk HPV, low-risk HPV, grouped 4-HPV-vaccine types, and grouped 9-HPV-vaccine-types, and the degree of concordance between the two tests was assessed. We also examined whether socio-demographic-behavioral factors were associated with concordance between the two assays. Results: The sensitivity of SPF10 PCR-DEIA-LiPA(25) was higher than Linear Array, with the exception of HPV 70, for the detection of oral HPV. The prevalence ratio of SPF10 PCR-DEIA-LiPA(25) to Linear Array varied between 1.0 and 9.0 for individual HPV genotypes, excluding HPV 70, and between 3.8 and 4.4 for grouped 4-valent and 9-valent HPV vaccine types, respectively. There was no association between socio-demographic-behavioral factors and discordance in results between the two tests for oral HPV 16 detection. Discussion: SPF10 PCR-DEIA-LiPA(25) was more sensitive than Linear Array for detecting HPV in oral gargle samples. Given the growing importance of detecting oral HPV infection for research studies of oral HPV natural history and vaccine effectiveness evaluation, we recommend using methods with higher sensitivity such as SPF10 PCR-DEIA-LiPA(25) for detecting HPV in oral gargle samples.