Construction of metal-organic framework-based multienzyme system for L-tert-leucine production.

Chiral compounds are important drug intermediates that play a critical role in human life. Herein, we report a facile method to prepare multi-enzyme nano-devices with high catalytic activity and stability. The self-assemble molecular binders SpyCatcher and SpyTag were fused with leucine dehydrogenase and glucose dehydrogenase to produce sc-LeuDH (SpyCatcher-fused leucine dehydrogenase) and GDH-st (SpyTag-fused glucose dehydrogenase), respectively. After assembling, the cross-linked enzymes LeuDH-GDH were formed. The crosslinking enzyme has good pH stability and temperature stability. The coenzyme cycle constant of LeuDH-GDH was always higher than that of free double enzymes. The yield of L-tert-leucine synthesis by LeuDH-GDH was 0.47 times higher than that by free LeuDH and GDH. To further improve the enzyme performance, the cross-linked LeuDH-GDH was immobilized on zeolite imidazolate framework-8 (ZIF-8) via bionic mineralization, forming LeuDH-GDH @ZIF-8. The created co-immobilized enzymes showed even better pH stability and temperature stability than the cross-linked enzymes, and LeuDH-GDH@ZIF-8 retains 70% relative conversion rate in the first four reuses. In addition, the yield of LeuDH-GDH@ZIF-8 was 0.62 times higher than that of LeuDH-GDH, and 1.38 times higher than that of free double enzyme system. This work provides a novel method for developing multi-enzyme nano-device, and the ease of operation of this method is appealing for the construction of other multi-enzymes @MOF systems for the applications in the kinds of complex environment.

Whole-genome sequencing and phylogenomic analyses of a novel zearalenone-degrading Bacillus subtilis B72.

Bacillus strain B72 was previously isolated as a novel zearalenone (ZEN) degradation strain from the oil field soil in Xinjiang, China. The genome of B72 was sequenced with a 400 bp paired-end using the Illumina HiSeq X Ten platform. De novo genome assembly was performed using SOAPdenovo2 assemblers. Phylogenetic analysis using 16S rRNA gene sequencing demonstrated that B72 is closely related to the novel Bacillus subtilis (B. subtilis) strain DSM 10. A phylogenetic tree based on 31 housekeeping genes, constructed with 19 strains closest at the species level, showed that B72 was closely related to B. subtilis 168, B. licheniformis PT-9, and B. tequilensis KCTC 13622. Detailed phylogenomic analysis using average nucleotide identity (ANI) and genome-to-genome distance calculator (GGDC) demonstrated that B72 might be classified as a novel B. subtilis strain. Our study demonstrated that B72 could degrade 100% of ZEN in minimal medium after 8 h of incubation, which makes it the fastest degrading strain to date. Moreover, we confirmed that ZEN degradation by B72 might involve degrading enzymes produced during the initial period of bacterial growth. Subsequently, functional genome annotation revealed that the laccase-encoding genes yfiH (gene 1743) and cotA (gene 2671) might be related to ZEN degradation in B72. The genome sequence of B. subtilis B72 reported here will provide a reference for genomic research on ZEN degradation in the field of food and feed. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03517-y.

Engineered living materials grown from programmable Aspergillus niger mycelial pellets.

The development of engineered living materials (ELMs) has recently attracted significant attention from researchers across multiple disciplines. Fungi-derived ELMs represent a new type of macroscale, cost-effective, environmentally sustainable materials. However, current fungi-based ELMs either have to undergo a final process to heat-kill the living cells or rely on the co-culture with a model organism for functional modification, which hinders the engineerability and versatility of these materials. In this study, we report a new type of ELMs - grown from programmable Aspergillus niger mycelial pellets - by a simple filtration step under ambient conditions. We demonstrate that A. Niger pellets can provide sufficient cohesion to maintain large-area self-supporting structures even under low pH conditions. Subsequently, by tuning the inducible expression of genes involved in melanin biosynthesis, we verified the fabrication of self-supporting living membrane materials with tunable colors in response to xylose concentration in the surroundings, which can be further explored as a potential biosensor for detecting xylose level in industrial wastewater. Notably, the living materials remain alive, self-regenerative, and functional even after 3-month storage. Thus, beyond reporting a new engineerable fungi chassis for constructing ELMs, our study provides new opportunities for developing bulk living materials for real-world applications such as the production of fabrics, packaging materials, and biosensors.

Recent advances in the construction of biocomposites based on fungal mycelia.

In recent years, environmental problems have become increasingly serious, significantly effecting the ecosystem and human health. To deal with the problem of environmental pollution in an eco-conscious way, sustainable composite biomaterials are being produced. Mycelium-based composite biomaterials combine biological systems with substrates such as nanomaterials or agricultural and industrial wastes, which can complement each other's advantages or turn waste into a useful resource. Such materials can solve practical wastewater problems as well as replace plastic products, thus reducing plastic pollution and contributing to the green transition of the environment. In this review, we summarized the recent findings of studies on these materials, indicating future research directions.