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Rao Ben,Liao Xianqing,Liu Fang,Chen Wei,Zhou Ronghua,Ma Lixin,Wang YaPing 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.6
L-amino acid oxidase (AAO) was reported to be capable of converting L-glutamic acid to α-aketoglutaric acid (α-KG). The sequence of AAO from Kitasatospora cheerisanensis was synthesized based on Pichia pastoris codon-usage preferences. AAO gene was cloned into plasmid pPICZα which was transformed into P. pastoris. Next, multi-copy expression plasmids were constructed by using plasmid pHBM905BDM. High-density fermentation was performed and the recombinant enzyme was characterized. The conversion conditions were optimized. By using Escherichia coli expression system, no soluble or active AAO was obtained from two strains after fermentation and induction. We can’t obtain high-level expression of recombinant strains by using plasmid pPICZα. Therefore, we constructed multi-copy expression plasmids using plasmid pHBM905BDM. By using this plasmid, multi-copy strains were constructed and named as PAAO1, PAAO2, PAAO3, PAAO4, and PAAO5, respectively. The following results showed that expression of AAO in multicopy strains increased as designed and strain PAAO5 was chosen for high-density fermentation and enzyme activity experiments. After high-density fermentation, we achieved an AAO-expression yield of 120.8 U/mL. After temperature and pH optimization, the highest AAO activity was observed at a temperature and pH of 20°C and 6, respectively. After optimization of the conversion conditions, the average production rate of L-glutamic acid to α-KG was 3.46 g/L/h and the highest α-KG titer (103 g/L) was converted from 120 g/L L-glutamic acid. In this study, AAO was abundantly expressed by using P. pastoris expression system. The following experiments indicated that AAO is suitable for use in industrial applications.
Impact of Multiple Satellite Ocean Color Samplings in a Day on Assessing Phytoplankton Dynamics
ZhongPing Lee,Mingshun Jiang,Curtiss Davis,Nima Pahlevan,안유환,Ronghua Ma 한국해양과학기술원 2012 Ocean science journal Vol.47 No.3
Ocean-color imagers on conventional polar-orbiting satellites have a revisit time of ~2 days for most regions, which is further reduced if the area is frequently cloudy. The Geostationary Ocean Color Imager (GOCI), the first ocean-color imager on a geostationary satellite, provides measurements 8times a day, thus significantly improving the frequency of measurements for studies of ocean environments. Here, we use results derived from GOCI measurements over Taihu Lake to demonstrate that the extra sampling can be used to improve the accuracy of statistically averaged longer-term (daily) measurements. Additionally, using numerical simulations, we demonstrate that the coupling of diurnal variations of both biomass and photosynthetic available radiation can improve the accuracy of daily primary production estimates. These results echo that higher sampling frequency can improve our estimates of longer-term dynamics of biogeochemical processes and highlights the value of ocean color measurements from geostationary satellites.