Discovery of a Fungal P450 with an Unusual Two-Step Mechanism for Constructing a Bicyclo[3.2.2]nonane Skeleton.

The successful biomimetic or chemoenzymatic synthesis of target natural products (NPs) and their derivatives relies on enzyme discovery. Herein, we discover a fungal P450 BTG5 that can catalyze the formation of a bicyclo[3.2.2]nonane structure through an unusual two-step mechanism of dimerization and cyclization in the biosynthesis of beticolin 1, whose bicyclo[3.2.2]nonane skeleton connects an anthraquinone moiety and a xanthone moiety. Further investigation reveals that BTG5-T318 not only determines the substrate selectivity but also alters the catalytic reactions, which allows the separation of the reaction to two individual steps, thereby understanding its catalytic mechanism. It reveals that the first heterodimerization undergoes the common oxidation process for P450s, while the second uncommon formal redox-neutral cyclization step is proved as a redox-mediated reaction, which has never been reported. Therefore, this work advances our understanding of P450-catalyzed reactions and paves the way for expansion of the diversity of this class of NPs through synthetic biology.

Synthetic Biology-based Construction of Unnatural Perylenequinones with Improved Photodynamic Anticancer Activities.

The construction of structural complexity and diversity of natural products is crucial for drug discovery and development. To overcome high dark toxicity and poor photostability of natural photosensitizer perylenequinones (PQs) for photodynamic therapy, herein, we aim to introduce the structural complexity and diversity to biosynthesize the desired unnatural PQs in fungus Cercospora through synthetic biology-based strategy. Thus, we first elucidate the intricate biosynthetic pathways of class B PQs and reveal how the branching enzymes create their structural complexity and diversity from a common ancestor. This enables the rational reprogramming of cercosporin biosynthetic pathway in Cercospora to generate diverse unnatural PQs without chemical modification. Among them, unnatural cercosporin A displays remarkably low dark toxicity and high photostability with retention of great photodynamic anticancer and antimicrobial activities. Moreover, it is found that, unlike cercosporin, unnatural cercosporin A could be selectively accumulated in cancer cells, providing potential targets for drug development. Therefore, this work provides a comprehensive foundation for preparing unnatural products with customized functions through synthetic biology-based strategies, thus facilitating drug discovery pipelines from nature.

Enhanced production of aspochalasin D through genetic engineering of Aspergillus flavipes.

Aspochalasin D (AD) belongs to the polyketide-amino acid hybrid natural products with anti-cancer, anti-bacterial, and anti-fouling bioactivities. However, the low production limits its further application. In this study, AD was separated and identified from Aspergillus flavipes 3.17641. Next, besides the optimization of culture conditions using a single-factor experiment and response surface methodology, metabolic engineering was employed to increase the AD production. It shows that the deletion of the shunt gene aspoA and overexpression of the pathway-specific regulator aspoG significantly improve the AD production. Its production reached to 812.1 mg/L under the optimized conditions, with 18.5-fold increase. Therefore, this study not only provides a general method for improving the production of similar natural products in other fungi, but also enables the further biological function development of AD in agriculture and pharmaceutical. KEY POINTS: • The Aspochalasin D (AD) production was improved by optimizing culture conditions. • The deletion of the shunt gene aspoA increased the AD production. • Overexpression of the pathway regulator aspoG further improved the AD production.

Discovery of the Biosynthetic Pathway of Beticolin 1 Reveals a Novel Non-Heme Iron-Dependent Oxygenase for Anthraquinone Ring Cleavage.

This study used light-mediated comparative transcriptomics to identify the biosynthetic gene cluster of beticolin 1 in Cercospora. It contains an anthraquinone moiety and an unusual halogenated xanthone moiety connected by a bicyclo[3.2.2]nonane. During elucidation of the biosynthetic pathway of beticolin 1, a novel non-heme iron oxygenase BTG13 responsible for anthraquinone ring cleavage was discovered. More importantly, the discovery of non-heme iron oxygenase BTG13 is well supported by experimental evidence: (i) crystal structure and the inductively coupled plasma mass spectrometry revealed that its reactive site is built by an atypical iron ion coordination, where the iron ion is uncommonly coordinated by four histidine residues, an unusual carboxylated-lysine (Kcx377) and water; (ii) Kcx377 is mediated by His58 and Thr299 to modulate the catalytic activity of BTG13. Therefore, we believed this study updates our knowledge of metalloenzymes.

Dynamically tunable directional subwavelength beam propagation based on photonic spin Hall effect in graphene-based hyperbolic metamaterials.

Photonic spin Hall effect (PSHE) of type II hyperbolic metamaterials is achieved due to near filed interference, which provides a way to decide the propagation direction of subwavelength beam. In this paper, we propose graphene-based hyperbolic metamaterials (GHMMs), which is composed of the alternating graphene/SiO2 multilayer. The numerical results show that when a dipole emitter is placed at the boundary of the GHMMs, the subwavelength beam with λ/40 full-with half maximum can be excited and propagates along the left or right channel, which is dependent on polarization handedness. In addition, we further demonstrate that the unidirectional propagation angle can be dynamically tuned by changing the external electric field bias applied to graphene.

Graphene-based hyperbolic metamaterials for a tunable subwavelength dark hollow beam.

Hyperbolic metamaterials have recently been widely investigated in nanophotonics systems. Here, we propose an alternating graphene/${{\rm SiO}_2}$SiO2 multilayer structure as an anisotropic medium with hyperbolic dispersion. When in-plane and out-of-plane effective permittivity is negative and positive, respectively, the incident beam (transverse magnetic polarization wave) can be split into two subwavelength beams, and a dark hollow beam can be achieved for circularly polarized incidence. Also, the size of the dark hollow beam can be tuned by changing the Fermi level. Our method is believed to be used as a tunable optical tweezer for controlling molecules.

A tunable THz absorber consisting of an elliptical graphene disk array.

Herein, we present an adjustable absorber consisting of a periodically patterned elliptical graphene disk array, which absorbs in the THz region. When a circularly polarized light beam illuminates this structure, its absorption spectrum displays two absorption peaks, which originate from the F-P resonance of the fundamental graphene edge plasmon mode along the major and minor axes of the elliptical graphene disk. The position of these two absorption peaks can be modulated by changing the Fermi level of graphene. Furthermore, both absorption bands can merge into one broadband by changing the length of the major and minor axes. The full width at half maximum (FWHM) of the broadband can reach up to 3.52 THz. In addition, by changing the incident elliptically polarized light, the peak ratio between the two absorption bands can also be tuned to convert the double-band absorption to single-band absorption.

Biosynthesis of Natural and Unnatural Perylenequinones for Drug Development.

A wide range of perylenequinones (PQs) with diverse structures and versatile bioactivities have long been isolated, positioning them as highly promising agents for photodynamic therapy (PDT). However, the lack of an efficient and cost-effective method to obtain these compounds and to introduce structural diversity and complexity currently hinders their further research and application. In this concept, we present a comprehensive overview of the advancements in the biosynthetic pathways of natural PQs based on their structural classification, and also summarize recent progress in the biosynthesis of natural PQs and derivatives. These pioneering efforts may pave the way for structure modification and large-scale bioproduction of natural and unnatural PQs through synthetic biology strategies to promote their drug development.

Arbitrarily polarized bound states in the continuum with twisted photonic crystal slabs.

Arbitrary polarized vortex beam induced by polarization singularity offers a new platform for both classical optics and quantum entanglement applications. Bound states in the continuum (BICs) have been demonstrated to be associated with topological charge and vortex polarization singularities in momentum space. For conventional symmetric photonic crystal slabs (PhCSs), BIC is enclosed by linearly polarized far fields with winding angle of 2π, which is unfavorable for high-capacity and multi-functionality integration-optics applications. Here, we show that by breaking σz-symmetry of the PhCS, asymmetry in upward and downward directions and arbitrarily polarized BIC can be realized with a bilayer-twisted PhCS. It exhibits elliptical polarization states with constant ellipticity angle at every point in momentum space within the vicinity of BIC. The topological nature of BIC reflects on the orientation angle of polarization state, with a topological charge of 1 for any value of ellipticity angle. Full coverage of Poincaré sphere (i.e., [Formula: see text] and [Formula: see text]) and higher-order Poincaré sphere can be realized by tailoring the twist angles. Our findings may open up new avenues for applications in structured light, quantum optics, and twistronics for photons.

Designing a cercosporin-bioinspired bifunctional algicide with flocculation and photocatalysis for efficiently controlling harmful cyanobacterial blooms.

Harmful cyanobacterial blooms (HCBs) are spreading in freshwater ecosystems worldwide, adversely affecting drinking water supplies, aquatic production, recreational and tourism activities. Therefore, the efficient and environmentally friendly method is still of interest to be developed to effectively control HCBs. Inspired by the excellent algicidal activity of cercosporin (CP), a novel metal-free algaecide SiO2@EDU@CP (EDU, N-ethyl-N'-(3-dimethylaminopropyl)urea) with flocculation and photoremoval functions, was successfully designed and prepared in one-step to simultaneously introduce CP and EDU on SiO2 nanoparticles. It could rapidly form algae flocs in 20 min with 97.1% flocculation rate, and remove Microcystis aeruginosa within 12 h with 91.0% algicidal rate under 23 W compact fluorescent light irradiation without any leaked CP detected. Additionally, odorant ß-cyclocitral and toxin microcystin-LR were both photodegraded after treatment of SiO2@EDU@CP. Further mechanistic studies showed that the introduction of EDU significantly reversed the zeta potential of SiO2-COOH to achieve the flocculation through neutral charge, and the photophysical characterization of SiO2@EDU@CP revealed the improved charge separation ability to generate reactive oxygen species. More importantly, the utility of SiO2@EDU@CP was well demonstrated by its effectiveness for algae from Taihu Lake under natural sunlight and inability to regrow after treatment. This study not only establishes a bifunctional algicide SiO2@EDU@CP to efficiently control HCBs, but also provides design possibilities to develop more novel and efficient algicides for the better control of practical HCBs.

Exploiting extraordinary topological optical forces at bound states in the continuum.

Polarization singularities and topological vortices in photonic crystal slabs centered at bound states in the continuum (BICs) can be attributed to zero amplitude of polarization vectors. We show that such topological features are also observed in optical forces within the vicinity of BIC, around which the force vectors wind in the momentum space. The topological force carries force topological charge and can be used for trapping and repelling nanoparticles. By tailoring asymmetry of the photonic crystal slab, topological force will contain spinning behavior and shifted force zeros, which can lead to three-dimensional asymmetric trapping. Several off-Γ BICs generate multiple force zeros with various force distribution patterns. Our findings introduce the concepts of topology to optical force around BICs and create opportunities to realize optical force vortices and enhanced reversible forces for manipulating nanoparticles and fluid flow.

Analysis of zwitterionic membrane fouling mechanism caused by HPAM in the presence of electrolytes.

Membrane fouling has always been a tough issue that is urgent to solve. Electrolytes which are prevalent in wastewater have a major influence on membrane fouling. Therefore, it is of great significance to understand the role and fouling mechanism of electrolytes in the membrane fouling process. In this work, the zwitterionic membrane is used to process hydrolyzed poly(acrylamide) (HPAM) with the addition of electrolytes (CaCl2, NaCl). Meanwhile, the effect of different electrolytes on the zwitterionic membrane fouling process by hydrolyzed poly(acrylamide) (HPAM) is systematically investigated. It was found that the flux recovery ratio (FRR) of the zwitterionic membrane is nearly 100% after treating HPAM with the addition of electrolytes. Therefore, molecular dynamics (MD) simulations were applied to illustrate the impact of electrolytes on the change of foulant structures and confirm the consequent effect of electrolytes on membrane fouling. According to the experiment and MD simulation results, it is found that the positive ion layer which exists between the HPAM and zwitterionic surface results in the excellent fouling resistance performance of the zwitterionic membrane. The zwitterionic membrane fouling mechanism is analyzed, which is helpful to the understanding of zwitterionic membrane fouling in high salinity wastewater.

Anisotropic Photonics Topological Transition in Hyperbolic Metamaterials Based on Black Phosphorus.

Based on in-plane anisotropy of black phosphorus (BP), anisotropic photonics topological transition (PTT) can be achieved by the proposed hyperbolic metamaterials structure, which is composed of alternating BP/SiO2 multilayer. Through effective medium theory and calculated iso-frequency contour, PTT can be found by carefully choosing the incident plane and other parameters. With the finite element method and transfer matrix method, a narrow angular optical transparency window with angular full width at half maximum of 1.32° exists at PTT. By changing the working wavelength, thickness of SiO2, or electron doping of black phosphorus, the incident plane of realizing PTT can be modulated, and anisotropic PTT is achieved.

Reassessment of the role of CaCO3 in n-butanol production from pretreated lignocellulosic biomass by Clostridium acetobutylicum.

In this study, the role of CaCO3 in n-butanol production was further investigated using corn straw hydrolysate (CSH) media by Clostridium acetobutylicum CICC 8016. CaCO3 addition stimulated sugars utilization and butanol production. Further study showed that calcium salts addition to CSH media led to the increase in Ca2+ concentration both intracellularly and extracellularly. Interestingly, without calcium salts addition, intracellular Ca2+ concentration in the synthetic P2 medium was much higher than that in the CSH medium despite the lower extracellular Ca2+ concentrations in the P2 medium. These results indicated that without additional calcium salts, Ca2+ uptake by C. acetobutylicum CICC 8016 in the CSH medium may be inhibited by non-sugar biomass degradation compounds, such as furans, phenolics and organic acids. Comparative proteomics analysis results showed that most enzymes involved in glycolysis, redox balance and amino acids metabolism were up-regulated with CaCO3 addition. This study provides further insights into the role of CaCO3 in n-butanol production using real biomass hydrolysate.