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At least 1,171 records · Page 65

On Optical Dipole Moment and Radiative Recombination Lifetime of Excitons in WSe 2

Optical dipole moment is the key parameter of optical transitions, as it directly determines the strength of light–matter interaction such as intrinsic radiative lifetime. However, experimental determination of these fundamental properties of excitons in monolayer WSe 2 is largely limited, because the commonly used measurement, such as (time-resolved) photoluminescence, is inherently difficult to probe the intrinsic properties. For example, dark states below bright exciton can change the photoluminescence emission rate by orders of magnitude and gives an “effective” radiative lifetime distinctive from the intrinsic one. On the other hand, such “effective” radiative lifetime becomes important itself because it describes how dark states affect exciton dynamics. Unfortunately, the “effective” radiative lifetime in monolayer WSe 2 is also not determined as it requires photoluminescence measurement with resonant excitation, which is technically difficult. These difficulties are overcome here to obtain both the “intrinsic” and “effective” radiative lifetime experimentally. A framework is developed to determine the dipole moment and “intrinsic” radiative lifetime of delocalized excitons in monolayer WSe 2 from the absorption measurements. In addition, the “effective” radiative lifetime in WSe 2 is obtained through time-resolved photoluminescence and absolute quantum-yield measurement at resonant excitation. These results provide helpful information for fundamental understanding of exciton light–matter interaction in WSe 2 .

36 MATERIALS SCIENCE↗

Thylakoid-integrated recombinant Hcf106 participates in the chloroplast twin arginine transport system

The chloroplast twin arginine transport (cpTat) system distinguishes itself as a protein transport pathway by translocating fully folded proteins, using the proton-motive force (PMF) as the sole source of energy. The cpTat pathway is evolutionarily conserved with the Tat pathway found in the plasma membrane of many prokaryotes. The cpTat (Escherichia coli) system uses three proteins, Tha4 (TatA),Hcf106 (TatB), and cpTatC (TatC), to form a transient translocase allowing the pas-sage of precursor proteins. Briefly, cpTatC and Hcf106, with Tha4, form the initial receptor complex responsible for precursor protein recognition and binding in an energy-independent manner, while a separate pool of Tha4 assembles with the pre-cursor-bound receptor complex in the presence the PMF. Analysis by blue-native polyacrylamide gel electrophoresis (BN-PAGE) shows that the receptor complex, in the absence of precursor, migrates near 700 kDa and contains cpTatC and Hcf106with little Tha4 remaining after detergent solubilization. To investigate the role thatHcf106 may play in receptor complex oligomerization and/or stability, systematic cysteine substitutions were made in positions from the N-terminal transmembrane domain to the end of the predicted amphipathic helix of the protein. BN-PAGE analysis allowed us to identify the locations of amino acids in Hcf106 that were critical for interacting with cpTatC. Oxidative cross-linking allowed us to map interactions of the transmembrane domain and amphipathic helix region of Hcf106. In addition, we showed that in vitro expressed, integrated Hcf106 can interact with the precursor signal peptide domain and imported cpTatC, strongly suggesting that a subpopulation of the integrated Hcf106 is participating in competent cpTat complexes.

59 BASIC BIOLOGICAL SCIENCES↗

Biotransformation of Isoeugenol into Vanillin Using Immobilized Recombinant Cells Containing Isoeugenol Monooxygenase Active Aggregates

For efficiently enhancing the activity of isoeugenol monooxygenase, a whole cell overproducing active aggregate IEM720-18A was successfully fabricated via the fusion of amphiphilic short peptide 18A (EWLKAFYEKVLEKLKELF) and isoeugenol monooxygenase and then efficiently expressed in E. coli BL21 (DE3). Such resulting bacteria, E. coli BL21 (DE3) harboring pET30a-IEM720-18A was applied in the biotransformation of isoeugenol to vanillin with the optimization of cultivation conditions. Our results revealed that the vanillin concentration reached to the highest level (14.5 mmol/L) under the optimized reaction conditions including 1.5-g cells containing active aggregate of IEM720-18A, 10% (v/v) dimethyl sulfoxide (DMSO), 100 mmol/L isoeugenol, 50 mmol/L glycine-sodium hydroxide buffer (pH 10.5) in 10 mL reaction volume, and 200 rpm at 25 °C for 36 h. Moreover, the active aggregate IEM720-18A was immobilized with 100 mmol/L glutaraldehyde at 4 °C to improve the operational stability. The highest activity could be achieved when the reactions were carried out at 25 °C and the relative activity of the immobilized enzyme maintained over 60% after seven recycles. Our study provides a new approach to the biotransformation of isoeugenol into vanillin.

60 APPLIED LIFE SCIENCES↗