Enhanced microwave metrology using an optical grating in Rydberg atoms.

An enhanced measurement of the microwave (MW) electric (E) field is proposed using an optical grating in Rydberg atoms. Electromagnetically induced transparency (EIT) of Rydberg atoms appears driven by a probe field and a control field. The EIT transmission spectrum is modulated by an optical grating. When a MW field drives the Rydberg transition, the central principal maximum of the grating spectrum splits. It is interesting to find that the magnitude of the sharp grating spectrum changes linearly with the MW E-field strength, which can be used to measure the MW E-field. The simulation result shows that the minimum detectable E-field strength is nearly 1/8 of that without gratings, and its measurement accuracy could be enhanced by about 60 times. Other discussion of MW metrology based on a grating spectrum is also presented.

Engineering the filamentous fungus Penicillium oxalicum for rapid, low-background and efficient protein expression.

Filamentous fungi are widely used in the field of recombinant protein expression due to their well-established protein modification systems and excellent secretion capacities. Although Penicillium oxalicum has been developed as an expression host, its potential for efficient and convenient protein production has not been fully exploited. In this study, we obtained an engineered strain by dominant activation of the G protein PGA3 using a point-mutation method based on the low extracellular background P. oxalicum host Δ13A-OamyR. This genetically modified strain, OamyR-QL, with faster cell growth and a more efficient Pamy15A promoter, will be used to construct a novel expression system. The relevant genes and pathways involved in the response to the G protein dominant activation in the engineered strain were revealed by RNA sequencing. Moreover, the transcription activator AmyR was overexpressed in OamyR-QL, resulting in a dramatically enhanced efficiency of the Pamy15A promoter. The construction of an efficient, low-background system by utilizing the G protein-AmyR regulatory pathway provides not only a theoretical reference for the genetic engineering of other filamentous fungal strains, but also a preferable option for the efficient and high purity expression of recombinant proteins in filamentous fungi.

Genetic modifications of critical regulators provide new insights into regulation modes of raw-starch-digesting enzyme expression in Penicillium.

BACKGROUND: Starch is a very abundant and renewable carbohydrate and an important feedstock for industrial applications. However, most starch-based products are not cost-efficient due to the high energy input needed in traditional enzymatic starch conversion processes. Raw-starch-digesting enzymes (RSDEs) from filamentous fungi have great commercial value in starch processing. However, the regulatory mechanisms associated with their production in filamentous fungi remain unknown. RESULTS: In this study, we reported the novel finding that cellulolytic fungus Penicillium oxalicum 114-2 has broad RSDE activity. Four regulators, including the amylase transcription activator AmyR, the catabolite repression repressor CreA, the group III G protein α subunit PGA3, and the nonhistone chromosomal protein HepA, have been found to play a crucial regulatory role in RSDE expression. Enzymatic assays revealed that RSDE production significantly increased after the overexpression of AmyR and HepA, the deletion of CreA and the dominant activation of PGA3. RT-qPCR analysis demonstrated that there is a mutual regulation mode between the four regulators, and then formed a cascade regulation mechanism that is involved in RSDE expression. Comparative transcriptomic analysis between the wild-type strain and genetically engineered strains revealed differentially expressed genes that may mediate the RSDE expression. CONCLUSIONS: The four different types of regulators were systematically investigated and found to form a regulatory network controlling RSDE gene expression. Our results provide a new insight into the regulatory mechanism of fungal amylolytic enzyme expression and offer a theoretical basis to rationally improve the RSDE yield in the future.

Construction of a novel filamentous fungal protein expression system based on redesigning of regulatory elements.

Filamentous fungi are extensively used as an important expression host for the production of a variety of essential industrial proteins. They have significant promise as an expression system for protein synthesis due to their inherent superior secretory capabilities. The purpose of this study was to develop a novel expression system by utilizing a Penicillium oxalicum strain that possesses a high capacity for protein secretion. The expression of glycoside hydrolases in P. oxalicum was evaluated in a cleaner extracellular background where the formation of two major amylases was inhibited. Four glycoside hydrolases (CBHI, Amy15B, BGL1, and Cel12A) were expressed under the highly constitutive promoter PubiD. It was found that the proteins exhibited high purity in the culture supernatant after cultivation with starch. Two inducible promoters, Pamy15A and PempA, under the activation of the transcription factor AmyR were used as elements in the construction of versatile vectors. When using the cellobiohydrolase CBHI as the extracellular quantitative reporter, the empA promoter screened from the AmyR-overexpressing strain was shown to be superior to the amy15A promoter based on RNA-sequencing data. Therefore, we designed an expression system consisting of a cleaner background host strain and an adjustable promoter. This system enables rapid and high-throughput evaluation of glycoside hydrolases from filamentous fungi.Key points• A new protein expression system derived from Penicillium oxalicum has been developed.• The expression platform is capable of secreting recombinant proteins with high purity.• The adjustable promoter may allow for further optimization of recombinant protein synthesis.

Design and Syntheses of Novel Fluoroporphyrin-Anthraquinone Complexes as Antitumor Agents.

A novel fluoroporphyrin-anthraquinone hybrid with dipeptide link and its metal complexes were synthesized and evaluated for anti-proliferation activity in human cancer cell line HeLa. The preliminary results demonstrated that all the compounds showed moderate to excellent antitumor activities. Among the active compounds, compound 3 which contains fluorinated porphyrin-anthraquinone and zinc ion exhibited the highest potency with IC50 value of 8.83 µM, indicating that it was a promising antitumor candidate.