Betulinic acid (BA) is a lupinane-type pentacyclic triterpenoid natural product derived from lupeol that has favorable anti-inflammatory and anti-tumor activities. Currently, BA is mainly produced via botanical extraction, which significantly limits its widespread use. In this study, we investigated the de novo synthesis of BA in Saccharomyces cerevisiae, and to facilitate the synthesis and storage of hydrophobic BA, we adopted a dual-engineering strategy involving peroxisomes and lipid droplets to construct the BA biosynthetic pathway. By expressing Betula platyphylla-derived lupeol C-28 oxidase (BPLO) and Arabidopsis-derived ATR1, we succeeded in developing a BA-producing strain and following multiple expression optimizations of the linker between BPLO and ATR1, the BA titer reached 77.53 mg/L in shake flasks and subsequently reached 205.74 mg/L via fed-batch fermentation in a 5-L bioreactor. In this study, we developed a feasible approach for the de novo synthesis of BA and its direct precursor lupeol in engineered S. cerevisiae.

Subcellular compartmentalization of metabolic pathways plays a crucial role in metabolic engineering. The peroxisome has emerged as a highly valuable and promising compartment for organelle engineering, particularly in the fields of biological manufacturing and agriculture. In this review, we summarize the remarkable achievements in peroxisome engineering in yeast, the industrially popular biomanufacturing chassis host, to produce various biocompounds. We also review progress in plant peroxisome engineering, a field that has already exhibited high potential in both biomanufacturing and agriculture. Moreover, we outline various experimentally validated strategies to improve the efficiency of engineered pathways in peroxisomes, as well as prospects of peroxisome engineering.

Bermudagrass is a perennial herb with the potential to remediate Pb pollution in soils, and it has mechanical resistance to shearing. However, the effects of mowing on Pb absorption and accumulation in bermudagrass are still unclear. In this study, we investigated the effects of different quantities (0, 1, 2, 4 applications) of mowing treatments under 200 mg L-1 Pb application on Pb accumulation and transport in bermudagrass and explored the related mechanisms. Compared to the Pb treatment, all of the mowing treatments greatly decreased root Pb concentration/accumulation, significantly enhanced Pb concentrations/accumulations in stubble stems and stubble leaves, and ultimately promoted Pb enrichment and transport. Of the treatments in this study, two applications of mowing best promoted Pb enrichment, and four applications of mowing best promoted Pb transport efficiency. Furthermore, mowing mediated the microdistribution and physiological patterns of Pb in bermudagrass and affected the Pb transport by changing the subcellar distribution patterns and chemical forms of Pb in various tissues. Additionally, mowing promoted the transport of all mineral elements and showed a synergistic relationship with Pb absorption and transport. The change in mineral element metabolism patterns may be an important reason why mowing promoted Pb accumulation in bermudagrass. Our study provides the first comprehensive evidence regarding mowing facilitating the absorption, accumulation and transport of Pb in bermudagrass. Moderate mowing may be an effective strategy to assist in soil Pb remediation using bermudagrass.

Lead (Pb) is one of the most harmful, toxic pollutants to the ecological environment and humans. Centipedegrass, a fast-growing warm-season turfgrass, is excellent for Pb pollution remediation. Exogenous low-molecular-weight organic acid (LMWOA) treatment is a promising approach for assisted phytoremediation. However, the effects of this treatment on the tolerance and Pb accumulation of centipedegrass are unclear. This study investigated these effects on the physiological growth response and Pb accumulation distribution characteristics of centipedegrass. Applications of 400 μM citric acid (CA), malic acid (MA) and tartaric acid (TA) significantly reduced membrane lipid peroxidation levels of leaves and improved biomass production of Pb-stressed plants. These treatments mainly increased peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) activities and enhanced free protein (Pro), ascorbic acid (AsA) and phytochelatins (PCs) contents, ultimately improving the Pb tolerance of centipedegrass. Their promoting effects decreased as follows: TA>CA>MA. All the treatments decreased root Pb concentrations and increased stem and leaf Pb concentrations, thus increasing total Pb accumulation and TF values. MA had the best and worst effects on Pb accumulation and Pb transportation, respectively. CA had the best and worst effects on Pb transportation and Pb accumulation, respectively. TA exhibited strong effects on both Pb accumulation and transport. Furthermore, all treatments changed the subcellular Pb distribution patterns and distribution models of the chemical forms of Pb in each tissue. The root Pb concentration was more highly correlated with the Pb subcellular fraction distribution pattern, while the stem and leaf Pb concentrations were more highly correlated with the distribution models of the chemical forms of Pb. Overall, TA improved plant Pb tolerance best and promoted both Pb absorption and transportation well and is considered the best candidate for Pb-contaminated soil remediation with centipedegrass. This study provides a new idea for Pb-contaminated soil remediation with centipedegrass combined with LMWOAs.

Dwarf bamboo Sasa argenteostriata (Regel) E.G. Camus is considered as potential plants for metal phytoremediation in previous filed observations. However, the mechanisms of lead (Pb) detoxification has not been described. The objective of this study was to explore the difference strategies or mechanisms of Pb detoxification in plant tissues. In this regard, four Pb treatments with hydroponics including 0 (control), 300, 600, and 900 mg L-1 were conducted to examine subcellular compartmentalization, Pb accumulation/species and antioxidant-assisted chelation. Our findings showed the retention of Pb by the whip-root system is one of its detoxification mechanisms to avoid damage the shoots. In addition, the cell wall retention is the dominant detoxification strategy of whips, new roots, old roots and new/old stems, while vacuolar compartmentalization is for new/old leaves. Interestingly, four low-mobility/-toxicity Pb species (i.e., FNaCl, FHAc, FHCl and FR) are distributed in roots, whips and stems, while two high-mobility/-toxicity Pb species (FE and FW) in leaves. The conversion of Pb to low-toxicity/-migration is a Pb-detoxification strategy in roots, whips and stems but not in leaves. Besides, the new/old roots and leaves can alleviate Pb damage through the synthesis of non-protein thiol, glutathione and phytochelatins. Among these, phytochelatins play a leading role in the detoxification in new/old roots, while glutathione is in new/old leaves. This study provides the first comprehensive evidence regarding the different strategies for Pb detoxification in dwarf bamboo tissues from physiological to cellular level, supporting that this plant could be rehabilitated for phytoremediation in Pb-contaminated media.

Despite not being an essential element for plants, Se has been proved to reduce Cd accumulation and Cd-induced oxidative stress, although the underlying mechanisms are not fully understood. A pak choi hydroponic experiment was conducted to investigate the effects of Se on Cd accumulation, subcellular distribution, and Cd-induced oxidative stress at different growth stages. The results showed that on day 19 after germination, Cd content was significantly reduced by 32% by selenite, but was increased by 15% by selenate. Accordingly, selenite improved cell-wall Cd sequestration by 20%, whereas selenate caused enhanced translocation of Cd from the root to the shoot. However, the effects of selenite on the reduction in Cd accumulation and distribution in pak choi seedlings were completely dismissed on day 40. Nevertheless, both forms of Se enhanced antioxidative defense, as they both inhibited the accumulation of H2O2 and malondialdehyde. On day 19, ascorbate peroxidase and glutathione reductase activities were increased by more than 50% by selenite; additionally, superoxide dismutase, catalase, and peroxidase activities increased by up to 86%, 63%, and 24%, respectively, in the presence of selenite, when compared to Cd treatment alone. Activities of most of the antioxidants remained significantly unaffected by both forms of Se on day 40. Consequently, selenite and selenate affected Cd accumulation in pak choi seedlings by altering Cd subcellular distribution and by enhancing antioxidative defense, but such effects depended on the Se forms applied and the growth stage as well.