Algae-Derived Bioactive Molecules for the Potential Treatment of SARS-CoV-2.

The recently emerged COVID-19 disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has adversely affected the whole world. As a significant public health threat, it has spread worldwide. Scientists and global health experts are collaborating to find and execute speedy diagnostics, robust and highly effective vaccines, and therapeutic techniques to tackle COVID-19. The ocean is an immense source of biologically active molecules and/or compounds with antiviral-associated biopharmaceutical and immunostimulatory attributes. Some specific algae-derived molecules can be used to produce antibodies and vaccines to treat the COVID-19 disease. Algae have successfully synthesized several metabolites as natural defense compounds that enable them to survive under extreme environments. Several algae-derived bioactive molecules and/or compounds can be used against many diseases, including microbial and viral infections. Moreover, some algae species can also improve immunity and suppress human viral activity. Therefore, they may be recommended for use as a preventive remedy against COVID-19. Considering the above critiques and unique attributes, herein, we aimed to systematically assess algae-derived, biologically active molecules that could be used against this disease by looking at their natural sources, mechanisms of action, and prior pharmacological uses. This review also serves as a starting point for this research area to accelerate the establishment of anti-SARS-CoV-2 bioproducts.

Improving Microbial Cell Factory Performance by Engineering SAM Availability.

Methylated natural products are widely spread in nature. S-Adenosyl-l-methionine (SAM) is the secondary abundant cofactor and the primary methyl donor, which confer natural products with structural and functional diversification. The increasing demand for SAM-dependent natural products (SdNPs) has motivated the development of microbial cell factories (MCFs) for sustainable and efficient SdNP production. Insufficient and unsustainable SAM availability hinders the improvement of SdNP MCF performance. From the perspective of developing MCF, this review summarized recent understanding of de novo SAM biosynthesis and its regulatory mechanism. SAM is just the methyl mediator but not the original methyl source. Effective and sustainable methyl source supply is critical for efficient SdNP production. We compared and discussed the innate and relatively less explored alternative methyl sources and identified the one involving cheap one-carbon compound as more promising. The SAM biosynthesis is synergistically regulated on multilevels and is tightly connected with ATP and NAD(P)H pools. We also covered the recent advancement of metabolic engineering in improving intracellular SAM availability and SdNP production. Dynamic regulation is a promising strategy to achieve accurate and dynamic fine-tuning of intracellular SAM pool size. Finally, we discussed the design and engineering constraints underlying construction of SAM-responsive genetic circuits and envisioned their future applications in developing SdNP MCFs.

A local or a stranger? Comparison of autochthonous vs. allochthonous microalgae potential for bioremediation of coal mine drainage water.

Coal mining endangers the environment by contaminating of soil, surface, and ground water with coal mine drainage water (CMW) polluted by heavy metals. Microalgal cultures, hyper-accumulators of heavy metals, represent a promising solution for CMW biotreatment. A bottleneck of this approach is the availability of microalgal strains that combine a large capacity for heavy metal biocapture with a high resilience to their toxic effects. Biotopes contaminated with heavy metals are frequently inhabited by microalgae evolved to be resilient to heavy metal toxicity. Therefore, the autochthonous (locally isolated) microalgal strains are a priori considered to be superior for biotreatment of heavy metal-polluted waste streams. Still, strains from biocollections combine a high pollutant resilience with other biotechnologically important traits such as high productivity, high CO2 sequestration rate etc. Moreover, the strains available "off-the-shelf" would enable rapid development of bioprocesses. Here, we compared the efficiency of CMW biotreatment with autochthonous (isolated from the coal mine drainage sump) and allochthonous microalgae (from a geographically distant phosphate-polluted site). Both autochthonous strains and allochthonous strains turned to be interchangeable under our experimental conditions. Still, the autochthonous strains showed a higher capacity for sequestration of iron, zinc, and manganese, the specific pollutants of the studied CMW. It can be important when the duration of unattended exploitation of the CMW treatment facility is a priority or spikes of the heavy metal concentration in CMW are expected. Therefore, the "off-the-shelf" strains can be a plausible solution for rapid development of CMW treatment technologies from scratch (although screening for acute toxicity of CMW is imperative). On the other hand, locally isolated strains can offer distinct advantages and should be always considered if sufficient time and other resources are available for the development of microalgae-based process for CMW treatment.

A novel integrated approach employing Desertifilum tharense BERC-3 for efficient wastewater valorization and recycling for developing peri-urban algae farming system.

Peri-urban environments are significant reservoirs of wastewater, and releasing this untreated wastewater from these resources poses severe environmental and ecological threats. Wastewater mitigation through sustainable approaches is an emerging area of interest. Algae offers a promising strategy for carbon-neutral valorization and recycling of urban wastewater. Aiming to provide a proof-of-concept for complete valorization and recycling of urban wastewater in a peri-urban environment in a closed loop system, a newly isolated biocrust-forming cyanobacterium Desertifilum tharense BERC-3 was evaluated. Here, the highest growth and lipids productivity were achieved in urban wastewater compared to BG11 and synthetic wastewater. D. tharense BERC-3 showed 60-95% resource recovery efficiency and decreased total dissolved solids, chemical oxygen demand, biological oxygen demand, nitrate nitrogen, ammonia nitrogen and total phosphorus contents of the water by 60.37%, 81.11%, 82.75%, 87.91%, 85.13%, 85.41%, 95.87%, respectively, making it fit for agriculture as per WHO's safety limits. Soil supplementation with 2% wastewater-cultivated algae as a soil amender, along with its irrigation with post-treated wastewater, improved the nitrogen content and microbial activity of the soil by 0.3-2.0-fold and 0.5-fold, respectively. Besides, the availability of phosphorus was also improved by 1.66-fold. The complete bioprocessing pipeline offered a complete biomass utilization. This study demonstrated the first proof-of-concept of integrating resource recovery and resource recycling using cyanobacteria to develop a peri-urban algae farming system. This can lead to establishing wastewater-driven algae cultivation systems as novel enterprises for rural migrants moving to urban areas.

Isolating Fistulifera pelliculosa from the northern Bohai Sea and analyzing biochemical composition, antibacterial and nutrient removal potential.

The microalgae located near the estuary of the Liaohe River along the coast of Panjin have long been in an area with large fluctuations in salinity, temperature, and nutrients, and have high-quality alternatives for high-value metabolites. Three strains of microalgae were screened and the biomass of microalgae could be optimized 0.313-0.790 g L-1 in 10 L bioreactor. The determination results of bioactive substances in these three microalgae showed that, the amount of fucoxanthin in the growth phase II (14 days) was maximum, at 5.354, 6.284 and 14.837 mg g-1 respectively. The diatoxanthin of Dut-wj-J1 in growth phase III (21 days) could reach 5.158 mg g-1. Dut-wj-J4 had the highest lipid production efficiency (9.45 mg L-1 d-1) followed by Dut-wj-J2 (8.49 mg L-1 d-1) and Dut-wj-J1 (8.18 mg L-1 d-1) respectively. These bioactive substances have inhibition zones of 7-13 mm against all four strains of bacteria ie., Acetobacter, Rhodococcus erythropolis, Escherichia coli and Bacillus subtilis Cohn respectively. In addition, these microalgae can play a potential role in nutrient enrichment in eutrophic seawater. The NO3- degradation rates of these three algae in the first 14 days were 75.0 %, 45.8 % and 100 % respectively, as well as the PO4- degradation rates in the first 7 days were 94.8 %, 100 % and 80.9 % respectively. This work manifests the plasticity of algae isolated from the Bohai Sea and provides useful insights for further joint production of bioactive substances.

Produce D-allulose from non-food biomass by integrating corn stalk hydrolysis with whole-cell catalysis.

D-allulose is a high-value rare sugar with many health benefits. D-allulose market demand increased dramatically after approved as generally recognized as safe (GRAS). The current studies are predominantly focusing on producing D-allulose from either D-glucose or D-fructose, which may compete foods against human. The corn stalk (CS) is one of the main agricultural waste biomass in the worldwide. Bioconversion is one of the promising approach to CS valorization, which is of significance for both food safety and reducing carbon emission. In this study, we tried to explore a non-food based route by integrating CS hydrolysis with D-allulose production. Firstly we developed an efficient Escherichia coli whole-cell catalyst to produce D-allulose from D-glucose. Next we hydrolyzed CS and achieved D-allulose production from the CS hydrolysate. Finally we immobilized the whole-cell catalyst by designing a microfluidic device. Process optimization improved D-allulose titer by 8.61 times, reaching 8.78 g/L from CS hydrolysate. With this method, 1 kg CS was finally converted to 48.87 g D-allulose. This study validated the feasibility of valorizing corn stalk by converting it to D-allulose.

Determination of active sites on the synthesis of novel Lewis acidic deep eutectic solvent catalysts and kinetic studies in microalgal biodiesel production.

Experimental and theoretical considerations for kinetic modeling of the transesterification reaction of microalgae lipids into biodiesel were investigated using Lewis acid deep eutectic solvents (DESs) as a catalyst. The acid sites involved in the reaction were characterized using acetonitrile as a probe to understand the mechanism. DES ChCl-SnCl2 (choline chloride-tin ii chloride) showed higher catalytic activity in transesterification due to its higher acidity compared to DES ChCl-ZnCl2 (choline chloride-zinc chloride). This was illustrated by geometric optimization of the DES structures through density functional theory (DFT) which showed that the metal centers furthest from the choline moiety are the most acidic and the bond lengths of Sn-Cl were between 2.56 and 2.77 Å, and were greater than the Zn-Cl bond lengths from 2.30 to 2.48 Å, making the ChCl-SnCl2 DES more acidic and more suitable for the biodiesel production. The fatty acid methyl ester (FAME) conversion from microalgae lipid was 36.75 mg g-1 under ideal conditions (6 molar ratio methanol-lipid with 8 vol% DES dosage in methanol at 140 °C for 420 min). The activation energy is found to be 36.3 kJ mol-1 based on the pseudo-first-order reaction, in addition, the DES catalyst (ChCl-SnCl2) drove the reaction chemically and did not show mass transfer limitation. Information from this study can help to advance the development of an efficient and environmentally friendly industrial biodiesel production technology.

Employing Cationic Kraft Lignin as Additive to Enhance Enzymatic Hydrolysis of Corn Stalk.

A water-soluble cationic kraft lignin (named JLQKL50), synthesized by combining quaternization and crosslinking reactions, was used as an additive to enhance the enzymatic hydrolysis of dilute-alkali-pretreated corn stalk. The chemical constitution of JLQKL50 was investigated by Fourier transform infrared spectroscopy, 1H nuclear magnetic resonance (NMR) and 13C NMR spectroscopy, and elemental analysis. The enzymatic hydrolysis efficiency of corn stalk at solid content of 10% (w/v) was significantly improved from 70.67% to 78.88% after 24 h when JLQKL50 was added at a concentration of 2 g/L. Meanwhile, the enzymatic hydrolysis efficiency after 72 h reached 91.11% with 10 FPU/g of cellulase and 97.92% with 15 FPU/g of cellulase. In addition, JLQKL50 was found capable of extending the pH and temperature ranges of enzymatic hydrolysis to maintain high efficiency (higher than 70%). The decrease in cellulase activity under vigorous stirring with the addition of JLQKL50 was 17.4%, which was much lower than that (29.7%) without JLQKL50. The addition of JLQKL50 reduced the nonproductive adsorption of cellulase on the lignin substrate and improved the longevity, dispersity, and stability of the cellulase by enabling electrostatic repulsion. Therefore, the enzymatic hydrolysis of the corn stalk was enhanced. This study paves the way for the design of sustainable lignin-based additives to boost the enzymatic hydrolysis of lignocellulosic biomass.

Engineering an efficient whole-cell catalyst for d-allulose production from glycerol.

d-Allulose has many health-benefiting properties and therefore sustainably applied in food, pharmaceutical, and nutrition industries. The aldol reaction-based route is a very promising alternative to Izumoring strategy in d-allulose production. Remarkable studies reported in the past cannot get rid of by-product formation and costly purified enzyme usage. In the present study, we explored the glycerol assimilation by modularly assembling the d-allulose synthetic cascade in Escherichia coli envelop. We achieved an efficient whole-cell catalyst that produces only d-allulose from cheap glycerol feedstock, eliminating the involvement of purified enzymes. Detailed process optimization improved the d-allulose titer by 1500.00%. Finally, the production was validated in 3-L scale using a 5-L fermenter, and 5.67 g L-1 d-allulose was produced with a molar yield of 31.43%.

Assessment of the Tolerance of a Chlorophyte Desmodesmus to CuO-NP for Evaluation of the Nanopollution Bioremediation Potential of This Microalga.

Broad application of CuO nanoparticles (CuO-NP) for industrial and household purposes leads to a continuous increase in their discharge to, and, hence, ever-increasing environmental hazards for aquatic ecosystems. Microalgae-based technologies hold promise for bioremediation of diverse hazardous micropollutants (HMP), including NP, from wastewater. In this study, we tested the ability of the green microalga Desmodesmus sp. to accumulate CuO-NP or their components. We also assessed the tolerance of this microalga to the environmentally relevant concentrations of CuO-NP. Using scanning electron microscopy, we demonstrated that the average size of CuO-NP was 50-100 nm, and their purity was confirmed with elemental composition analysis. Tests of the colloidal suspensions of CuO-NP showed that the hydrodynamic diameter of CuO-NP and their aggregates was below 100 nm. Flow cytometry analysis showed that CuO-NP at a concentration of 100 µg L-1 slightly inhibited the viability of microalgae cells and led to an increase in their oxidative stress. The assessment of the condition of photosystem II showed that CuO-NP exert a multifaceted effect on the photosynthetic apparatus of Desmodesmus sp., depending on the concentration of and the exposure to the CuO-NP. Desmodesmus sp. turned to be relatively tolerant to CuO-NP. In addition, the ICP-MS method revealed increased bioaccumulation of copper by microalgae cells in the experimental groups. The outcomes of this study indicate that the Desmodesmus sp. has a significant potential for bioremoval of the copper-based nanostructured HMP from an aquatic environment.

Recent advances in photochemical and electrochemically induced thiocyanation: a greener approach for SCN-containing compound formation.

Techniques utilizing photo- and electrochemically induced reactions have been developed to accelerate organic processes. These techniques use light or electrical energy (electron transfer) as a direct energy source without using an initiator or reagent. Thiocyanates are found in biologically active and pharmacological compounds and can be converted into various functional groups. It is one of the most prominent organic scaffolds. Significant development in photo- and electro-chemically induced thiocyanation procedures has been made in recent years for the conception of carbon-sulfur bonds and synthesis of pharmaceutically important molecules. This review discusses different photo- and electro-chemically driven thiocyanation C(sp3)-SCN, C(sp2)-SCN, and C(sp)-SCN bond conception processes that may be useful to green organothiocyanate synthesis. We focus on the synthetic and mechanistic characteristics of organic photo- and electrochemically accelerated C-SCN bond formation thiocyanation reactions to highlight major advances in this novel green and sustainable research field.

Prevalence of anxiety, depression, and stress among students of Jahangirnagar University in Bangladesh.

Background and Aims: Anxiety and depression, as well as stress, are well-known problems observed across the world, particularly among students. This study intends to identify the level of anxiety, depression, and stress among university students and determine its association with their sociodemographic characteristics. Methods: The primary data were collected from 351 students with the help of a self-administrated questionnaire consisting of sociodemographic information and contains the Depression, Anxiety and Stress Scale-21 Items (DASS-21) instrument over the period December 8, 2019-January 23, 2020. The χ 2 test is employed to find the association between the status of stress, anxiety, and depression level with selected sociodemographic variables, and confirmatory factor analysis is used to find interrelationships between DASS items. Results: The results illustrate that no students have an extremely severe level of stress. However, the majority have a mild or moderate level of stress and it is associated with sex and residence (urban or rural). More than 40% of students have extremely severe anxiety. Results reveal that gender, residence (urban or rural), and family type of the students are linked with the anxiety level. The results also suggest that the type of accommodation of the students, their family type, and birth order are related to their depression level at a 5% level of significance. The findings also disclose that female students have more levels of depression, stress, and anxiety than their counterparts. Conclusion: Considering the finding, the authors think that the university authority should pay a need for greater interest to the mental well-being of students to enhance their quality of life. Given the harmful impacts of stress on academic performance and health, university administrators should be incorporating anxiety, stress, and depression management training in orientation activities.

Coupling with in-situ electrochemical reactive chlorine species generation and two-phase partitioning method for enhanced microalgal biodiesel production.

Microalgal-based biofuel production is of great significance in alleviating energy crisis and achieving carbon neutrality. However, the excessive costs and high solvent consumption in lipids extraction from microalgal obstruct the widespread application of biodiesel in practice. Reported herein is the construction of facile strategy for lipids extraction via electrocatalytic pretreatment and a subsequent two-phase partitioning method. Electrocatalytic pretreatment method adopts the solar as power source and avoids the drying of microalgal biomass, in favor of carbon neutrality requirement. During this process, eco-friendly electrode with high specific surface area could contribute to the sufficient generation of reactive chlorine species (RCS), facilitating the outflows of intracellular lipid. As a result, assisted with two-phase partitioning method, a satisfied performance of lipid recovery (86.72 %) was obtained. Notably, compared with traditional solvent method, two-phase partitioning method greatly reduced the dosage of organic solvent, which is an economical or environmental technique.

Progress in biohythane production from microalgae-wastewater sludge co-digestion: An integrated biorefinery approach.

Recent advances in microalgae to biohythane (bio-H2 and bio-CH4) conversion have achieved growing attention due to their eco-friendly and energy-efficient nature. Although microalgae are considered a potential 3rd - 4th generation biomass, their low C/N ratio and cell-wall biopolymers are challenging for biohythane production. This study emphasizes the solutions to mitigate the adverse effects of microalgae-based biohythane production using co-digestion with wastewater sludge. Wastewater sludge, an emerging environmental concern, is reviewed to be an effective co-substrate with microalgae to establish a biorefinery approach. The future trends and prospects of this biorefinery approach is critically reviewed to attain a profitable process. This study also reviewed the advantages of microalgae-wastewater co-cultivation and the application of activated sludge for bio-flocculation as a cost-effective solution for microalgae cultivation and harvesting. Microalgae-wastewater co-cultivation is also recommended to be effective for biohythane purification. The liquid digestate is suggested to be used as a culture media to enhance microalgal growth; whereas, the solid digestate could be transformed into resources through hydrothermal processes as a solution of digestate management. A practical biorefinery approach combining the synergistic benefits of microalgae-wastewater sludge and its biological conversion to biohythane would be an adjoining link to the beginning of a sustainable future.

Prospects of algae-based green synthesis of nanoparticles for environmental applications.

Green synthesis of nanoparticles (NPs) has emerged as an eco-friendly alternative to produce nanomaterials with diverse physical, chemical, and biological characteristics. Previously used, physical and chemical methods involve the production of toxic byproducts, costly instrumentation, and energy-intensive experimental processes thereby, limiting their applicability. Biogenic synthesis of nanoparticles has come forward as a potential alternative, providing an eco-friendly, cost-effective, and energy-efficient approach for the synthesis of a diverse range of NPs. Several biological entities are employed in the biosynthesis of NPs including bacteria, fungi, and algae. However, the distinguishing characteristics of microalgae and cyanobacteria make them promising candidates for NPs synthesis because of their higher growth rate, substantially higher rate of sequestering CO2, hyperaccumulation of heavy metals, absence of toxic byproducts, minimum energy input, and employment of biomolecules (pigments and enzymes) as reducing and capping agents. Algal extract, being a natural reducing and capping agent, serves as a living cell factory for the efficient green synthesis of nanoparticles. Physiological and biological methods allow algal cells to uptake heavy metals and utilize them as nutrient source to generate biomass by regulating their metabolic processes. Despite their enormous potential, studies on the microalgae-based synthesis of nanoparticles for the removal of toxic pollutants from wastewater remained an unexplored research area in the literature. This review was aimed to summarize the recent advancements and prospects in the algae-based synthesis of nanoparticles for environmental applications particularly treating the wastewater.

Novel biorefining method for succinic acid processed from sugarcane bagasse.

Sugarcane bagasse (SCB) was pretreated with hot water (HLW), ethanol (ETH), and sodium hydroxide (SH). The obtained residuals were hydrolyzed and applied as carbon sources for succinic acid (SA) fermentation, the residue digestibility and SA conversion rate of alkali-pretreated residual were superior to others. Considering the characteristics of alkali pretreatment, enzymatic hydrolysis and succinic acid fermentation, a novel in-situ semi-simultaneous saccharification and co-fermentation (SSSCF) procedure for SA production from SCB was developed. The yield, productivity, and conversion rates of SA from SCB raw material (DRM) processed by SSSCF were 41 g/L, 300 mg/L/h, and 320 mg/g dry, respectively. For every kilogram of SA production, the developed coupling method reduced the SH and water usages, energy consumption, and effluent emission by 0.14 kg, 233.5 L 14,000 kJ and 7 L, respectively, and enhanced the SA productivity by 1.7 times compared with the non-coupling procedure.

Co-fermentation of succinic acid and ethanol from sugarcane bagasse based on full hexose and pentose utilization and carbon dioxide reduction.

The full utilization of carbohydrates in lignocellulosic biomass is essential for an efficient biorefining process. In this study, co-fermentation was performed for processing ethanol and succinic from sugarcane bagasse. By optimizing the co-fermentation conditions, nutrition and feeding strategies, a novel process was developed to make full utilization of the glucose and xylose in the hydrolysate of sugarcane bagasse. The achieved concentrations of succinic acid and ethanol reached to 22.1 and 22.0 g/L, respectively, and could realize the conversion of 100 g SCB raw material into 8.6 g ethanol and 8.7 g succinic acid. It is worth mentioning that the CO2 released from S. cerevisiae in co-fermentation system was recycled by A. succinogenes to synthesize succinic acid, realized CO2 emission reduction in the process of lignocellulosic biomass biorefinery. This study provided a clue for efficient biorefinery of lignocellulosic biomass and reduction greenhouse gas emissions.

Recent Advances in Producing Sugar Alcohols and Functional Sugars by Engineering Yarrowia lipolytica.

The sugar alcohols and functional sugars have wide applications in food, pharmaceutical, and chemical industries. However, the smaller quantities of natural occurring sugar alcohols and functional sugars restricted their applications. The enzymatic and whole-cell catalyst production is emerging as the predominant alternatives. The properties of Yarrowia lipolytica make it a promising sugar alcohol and functional sugar producer. However, there are still some issues to be resolved. As there exist reviews about the chemical structures, physicochemical properties, biological functions, applications, and biosynthesis of sugar alcohols and/or functional sugars in Y. lipolytica, this mini review will not only update the recent advances in enzymatic and microbial production of sugar alcohols (erythritol, D-threitol, and xylitol) and functional sugars (isomaltulose, trehalose, fructo-oligosaccharides, and galacto-oligosaccharides) by using recombinant Y. lipolytica but also focus on the studies of gene discovery, pathway engineering, expanding substrate scope, bioprocess engineering, and novel breeding methods to resolve the aforementioned issues.

Al-doped α-MnO2 coated by lignin for high-performance rechargeable aqueous zinc-ion batteries.

Zn/MnO2 batteries, one of the most widely studied rechargeable aqueous zinc-ion batteries, suffer from poor cyclability because the structure of MnO2 is labile with cycling. Herein, the structural stability of α-MnO2 is enhanced by simultaneous Al3+ doping and lignin coating during the formation of α-MnO2 crystals in a hydrothermal process. Al3+ enters the [MnO6] octahedron accompanied by producing oxygen vacancies, and lignin further stabilizes the doped Al3+ via strong interaction in the prepared material, Al-doped α-MnO2 coated by lignin (L + Al@α-MnO2). Meanwhile, the conductivity of L + Al@α-MnO2 improves due to Al3+ doping, and the surface area of L + Al@α-MnO2 increases because of the production of nanorod structures after Al3+ doping and lignin coating. Compared with the reference α-MnO2 cathode, the L + Al@α-MnO2 cathode achieves superior performance with durably high reversible capacity (∼180 mA h g-1 at 1.5 A g-1) and good cycle stability. In addition, ex situ X-ray diffraction characterization of the cathode at different voltages in the first cycle is employed to study the related mechanism on improving battery performance. This study may provide ideas of designing advanced cathode materials for other aqueous metal-ion batteries.

Mycolicibacterium cell factory for the production of steroid-based drug intermediates.

Steroid-based drugs have been developed as the second largest medical category in pharmaceutics. The well-established route of steroid industry includes two steps: the conversion of natural products with a steroid framework to steroid-based drug intermediates and the synthesis of varied steroid-based drugs from steroid-based drug intermediates. The biosynthesis of steroid-based drug intermediates from phytosterols by Mycolicibacterium cell factories bypasses the potential undersupply of diosgenin in the traditional steroid chemical industry. Moreover, the biosynthesis route shows advantages on multiple steroid-based drug intermediate products, more ecofriendly processes, and consecutive reactions carried out in one operation step and in one pot. Androsta-4-ene-3,17-dione (AD), androsta-1,4-diene-3,17-dione (ADD) and 9-hydroxyandrostra-4-ene-3,17-dione (9-OH-AD) are the representative steroid-based drug intermediates synthesized by mycolicibacteria. Other steroid metabolites of mycolicibacteria, like 4-androstene-17ß-ol-3-one (TS), 22-hydroxy-23,24-bisnorchol-4-ene-3-one (4-HBC), 22-hydroxy-23,24-bisnorchol-1,4-diene-3-one (1,4-HBC), 9,22-dihydroxy-23,24-bisnorchol-4-ene-3-one (9-OH-HBC), 3aα-H-4α-(3'-propionic acid)-7aß-methylhexahydro-1,5-indanedione (HIP) and 3aα-H-4α-(3'-propionic acid)-5α-hydroxy-7aß-methylhexahydro-1-indanone-δ-lactone (HIL), also show values as steroid-based drug intermediates. To improve the bio-production efficiency of the steroid-based drug intermediates, mycolicibacterial strains and biotransformation processes have been continuously studied in the past decades. Many mycolicibacteria that accumulate steroid drug intermediates have been isolated, and subsequently optimized by conventional mutagenesis and genetic engineering. Especially, with the clarification of the mycolicibacterial steroid metabolic pathway and the developments on gene editing technologies, rational design is becoming an important measure for the construction and optimization of engineered mycolicibacteria strains that produce steroid-based drug intermediates. Hence, by reviewing researches in the past two decades, this article updates the overall process of steroid metabolism in mycolicibacteria and provides comprehensive schemes for the rational construction of mycolicibacterial strains that accumulate steroid-based drug intermediates. In addition, the special strategies for the bioconversion of highly hydrophobic steroid in aqueous media are discussed as well.