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Microbial immobilization technology is considered an efficient bioremediation method for chromium (Cr) pollution. However, it is currently unclear which strain is more beneficial for the remediation of Cr-contaminated water and soil. Therefore, corn straw biochar was used as a carrier to prepare materials for fixing the endophytes Serratia sp. Y-13 (BSR1), Serratia nematodiphila (BSR2), Lysinibacillus sp. strain SePC-36 (BLB1), Lysinibacillus mangiferihumi strain WK63 (BLB2) and the commercial bacteria Shewanella oneidensis MR-1 (BSW). The results demonstrated that, compared with BSW, endophyte-loaded biochar (especially BSR1) was more effective at remediating Cr pollution in water and soil. Endophyte-loaded biochar reduced the abundance of soil pathogenic bacteria, enhanced the number of beneficial plant endophytes, reduced the soil Cr(VI) concentration, improved soil fertility, reduced the plant Cr concentration and improved the yield of lettuce. Redundancy analysis (RDA) and structural equation modelling (PLS-PM) suggested that soil microbes are closely related to soil Cr(VI), plant fresh weight and soil organic matter, whereas endophyte-loaded biochar directly influences plant cell motility pathways by altering plant microbes. This study represents a pioneering investigation into the efficacy of endophyte-loaded biochar as a remediation strategy for Cr pollution.

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Diosgenin is an important raw material used in the synthesis of steroid drugs, and it is widely used in the pharmaceutical industry. The traditional method of producing diosgenin is through using raw materials provided via the plant Dioscorea zingiberensis C. H. Wright (DZW), which is subsequently industrially hydrolyzed using a high quantity of hydrochloric and sulfuric acids at temperatures ranging from 70 °C to 175 °C. This process results in a significant amount of unmanageable wastewater, creates issues of severe environmental pollution and consumes high quantities of energy. As an alternative, the enzymolysis of DZW to produce diosgenin is an environmentally and friendly method with wide-ranging prospects for its application. However, there are still only a few enzymes that are suitable for production on an industrial scale. In this study, three new key enzymes, E1, E2, and E3, with a high conversion stability of diosgenin, were isolated and identified using an enzyme-linked-substrate autography strategy. HPLC-MS/MS identification showed that E1, a 134.45 kDa protein with 1019 amino acids (AAs), is a zinc-dependent protein similar to the M16 family. E2, a 97.89 kDa protein with 910 AAs, is a type of endo-ß-1,3-glucanase. E3, a 51.6 kDa protein with 476 AAs, is a type of Xaa-Pro aminopeptidase. In addition, the method to immobilize these proteins was optimized, and stability was achieved. The results show that the optimal immobilization parameters are 3.5% sodium alginate, 3.45% calcium chloride concentration, 1.4 h fixed time, and pH 8.8; and the recovery rate of enzyme activity can reach 43.98%. A level of 70.3% relative enzyme activity can be obtained after employing six cycles of the optimized technology. Compared with free enzymes, immobilized enzymes have improved stability, acid and alkaline resistance and reusability, which are conducive to large-scale industrial production.

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Sulfur autotrophic denitrification coupled anaerobic ammonia oxidation (SAD/A) has several advantages over other denitrification processes; for example, it does not consume the organic carbon source, has low operation costs, and produces less excess sludge; however, it has certain disadvantages as well, such as a long start-up time, easy loss of bacteria, and low microbial activity at low temperature. The use of microbial immobilization technology to embed functional bacteria provides a feasible method of resolving the above problems. In this study polyvinyl alcohol­sodium alginate was used to prepare a composite carrier for fixing anaerobic ammonia oxidizing bacteria (AAOB) and sulfur oxidizing bacteria (SOB), and the structure and morphology of the encapsulated bodies were characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. Subsequently, the nitrogen removal performance of the immobilized microbial carriers in the gradient cooling process (30 °C to 10 °C) was determined, and the corresponding mechanism was discussed. The results showed that the nitrate-removal efficiencies observed with granular sludge and gel embedding were at 10 °C 21.44 % and 14.31 % lower, than those at 30 °C, respectively, whereas the ammonia-removal efficiency decreased by up to approximately three-fold. The main mechanism was the 'insulation' provided by the external gel composed of PVA and SA for the internal sludge and subsequent improvement of its low temperature resistance, while protecting AAOB and SOB from oxygen inhibition, which is conducive to enriching denitrifying bacteria. In addition, the gel does not change the internal sludge species, it can shift the dominance of specific microorganisms and improve the removal efficiency of nitrogen. In summary, the immobilization of AAOB and SOB by the gel can achieve effectively mitigate nitrogen pollution in low temperature environments, thus indicating that the SAD/A process has broad engineering application prospects.

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The discharge of chromium (Cr) contaminated wastewater is creating a serious threat to aquatic environment due to the rapid pace in agricultural and industrial activities. Particularly, the long-term exposure of Cr(VI) polluted wastewater to the environment is causing serious harm to human health. Therefore, the treatment of Cr(VI) contaminated wastewater is demanding widespread attention. Regarding this, the bioremediation is being considered as a reliable and feasible option to handle Cr(VI) contaminated wastewater because of having low technical investment and operating costs. However, certain factors such as loss of microorganisms, toxicity to microorganisms and uneven microbial growth cycle in the presence of high concentrations of Cr(VI) are hindering its commercial applications. Regarding this, microbial immobilization technology (MIT) is getting great research interest because it could overcome the shortcomings of bioremediation technology's poor tolerance against Cr. Therefore, this review is the first attempt to emphases recent research developments in the remediation of Cr(VI) contamination via MIT. Starting from the selection of immobilized carrier, the present review is designed to critically discuss the various microbial immobilizing methods i.e., adsorption, embedding, covalent binding and medium interception. Further, the mechanism of Cr(VI) removal by immobilized microorganism has also been explored, precisely. In addition, three kinds of microorganism immobilization devices have been critically examined. Finally, knowledge gaps/key challenges and future perspectives are also discussed that would be helpful for the experts in improving MIT for the remediation of Cr(VI) contamination.

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BACKGROUND: Butyl acetate has shown wide applications in food, cosmetics, medicine, and biofuel sectors. These short-chain fatty acid esters can be produced by either chemical or biological synthetic process with corresponding alcohols and acids. Currently, biosynthesis of short chain fatty acid esters, such as butyl butyrate, through microbial fermentation systems has been achieved; however, few studies regarding biosynthesis of butyl acetate were reported. RESULTS: In this study, three proof-of-principle strategies for the one-pot butyl acetate production from glucose through microbial fermentation were designed and evaluated. (1) 7.3 g/L of butyl acetate was synthesized by butanol-producing Clostridium acetobutylicum NJ4 with the supplementation of exogenous acetic acid; (2) With the addition of butanol, 5.76 g/L of butyl acetate can be synthesized by acetate-producing Actinobacillus succinogenes130z (ΔpflA); (3) Microbial co-culture of C. acetobutylicum NJ4 and A. succinogenes130z (ΔpflA) can directly produce 2.2 g/L of butyl acetate from glucose by using microbial co-culture system with the elimination of precursors. Through the further immobilization of A. succinogenes130z (ΔpflA), butyl acetate production was improved to 2.86 g/L. CONCLUSION: Different microbial mono- and co-culture systems for butyl acetate biosynthesis were successfully constructed. These strategies may be extended to the biosynthesis of a wide range of esters, especially to some longer chain ones.

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ß-1,3-xylanase plays an important role in the conversion of algal biomass into renewable chemical commodities and functional xylooligosaccharides. Efficient immobilization of the target enzymes that could integrate the separation, purification, and immobilization in one-step may have great potentials. The cheap and easily obtained pitch was adopted as the raw material to prepare the hyper-cross-linked polymers with the specific surface areas of 560 m2/g and the Ni2+ loading about 80 mg/g. As a carrier, it could immobilize and purify the ß-1,3-xylanases directly from the crude cell debris with the activity recovery of (80.66 ± 0.87) %. The half-life of the immobilized ß-1,3-xylanases was 50 min at 50 °C, which was significantly longer than the free ones with only 14 min. The immobilized ß-1,3-xylanases could retain 65 % of the activity after stored at 30 ℃ for 80 h, they could also retain 87 % of the original activity after 15 cycles of reuse. In terms of potential applications in the manipulation of algae Caulerpa lentillifera, the immobilized ß-1,3-xylanases could release 30 % more reducing sugar in 85 h. The method proposed here is promising for algal biomass manipulation because of its low cost, stable storage, convenient recycling and excellent reusability.

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The continually growing use of glyphosate and its critically discussed health and biodiversity risks ask for fast, low cost, on-site sensing technologies for food and water. To address this problem, we designed a highly sensitive sensor built on the remarkably specific recognition of glyphosate by its physiological target enzyme 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPs). This principle is implemented in an interferometric sensor by using the recently established soft colloidal probe (SCP) technique. EPSPs was site-specifically immobilized on a transparent surface utilizing the self-assembling properties of circadian clock gene 2 hydrophobin chimera and homogeneity of the layer was evidenced by atomic force microscopy. Exposure of the enzyme decorated biochip to glyphosate containing samples causes formation of enzyme-analyte complexes and a competitive loss of available binding sites for glyphosate-functionalized poly(ethylene glycol) SCPs. Functionalization of the SCPs with different types of linker molecules and glyphosate was assessed employing confocal laser scanning microscopy as well as confocal Raman microspectroscopy. Overall, reflection interference contrast microscopy analysis of SCP-biochip interactions revealed a strong influence of linker length and glyphosate coupling position on the sensitivity of the sensor. In employing a combination of pentaglycine linker and tethering glyphosate via its secondary amino group, concentrations in aqueous solutions down to 100 pM could be measured by the differential adhesion between SCP and biochip surface, supported by automated image analysis algorithms. This sensing concept could even prove its exceptional pM sensitivity in combination with a superior discrimination against structurally related compounds.

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A novel microorganism embedding material was developed to enhance the benzene removal through adsorption and biodegradation, by introducing ß-cyclodextrin (CD) to traditional polyvinyl alcohol gel beads. Results show that the optimal ratio of sucrose/benzene was 1.25 for co-metabolism biodegradation of benzene, and the maximum exogenous microbial respiration rate was 260.13 mgO2/(gVSS h) for gel beads with CD. The positive effects of CD on benzene removal mainly resulted from the adsorption characteristics of CD as well as the stimulation of CD on microbial activity. Adsorption tests indicate that CD addition increased the adsorption function of gel beads to benzene with its dispersion coefficient of 5.1 × 10-7 cm2/s. Respiration tests show that gel beads with CD possessed the highest maximum specific exogenous respiration rates. Moreover, a high-throughput sequencing analysis confirms that CD addition could obviously enhance microbial diversity with domain microbial of Zoogloea (17.0%). Finally, microbial embedding gel beads could remove certain benzene after lyophilization and storage for one month. Overall, the novel microbial embedding gel beads modified with CD (a favorable additional agent to traditional embedding materials) have been proved as an efficient method for removing benzene under suitable sucrose/benzene ratio.

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Nitrification and anaerobic ammonia oxidation (ANAMMOX) sludge was acclimated via SBR and MBR reactors respectively, both of which were embedded with microbial immobilization technology and spliced together to construct a PN/A double-bacteria-layer system. A short-term experiment proved that ammonia-oxidizing bacteria (AOB) and ANAMMOX bacteria in the system played dominant roles separately in different stages, which maintained the acid-base balance and achieved efficient removal of NH4+-N (98.8%). A long-term experiment showed that the PN/A double-bacteria-layer system improved the utilization of dissolved oxygen (DO) and enhanced the stability and nitrogen removal efficiency, with limited dissolved oxygen. The nitrogen removal efficiency of the PN/A double-bacteria-layer system remained stable at~80% when the influent NH4+-N was 200 mg·L-1 or 400 mg·L-1 in the presence of 1.0 mg·L-1 DO, whereas those of the control group were only 58.1% and 61.4%, respectively. When influent NH4+-N and DO were 400 mg·L-1 and 3.0 mg·L-1, respectively, the PN/A double-bacteria-layer system achieved a nitrogen removal efficiency, nitrogen loading rate (NLR), and nitrogen removal rate (NRR) of 87.9%, 0.4 kg·(m3·d)-1, and 12.8 mg·(g·h)-1, respectively.

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This study investigated the immobilizations with of bacteria two kinds of algal materials, Enteromorpha residue and kelp residue. The lipophilicity of them were compared by diesel absorption rates. The immobilization efficiency of Bacillus sp. E3 was measured to evaluate whether these carriers would satisfy the requirement for biodegradation of oil spills. The bacteria were immobilized through adsorption with the sterilized and non-sterilized carriers to compare the differences between the two treatments. Oil degradation rates were determined using gravimetric and GC-MS methods. Results showed the absorption rates of Enteromorpha residue and kelp residue for diesel were 411 and 273% respectively and remained approximately 105 and 120% after 2 h of erosion in simulated seawater system. After immobilized of Bacillus sp. E3, the oil degradation rates of them were higher than 65% after 21 days biodegradations. GC-MS analysis showed that two immobilizations degraded higher than 70% of the total alkane and the total PAHs, whereas the free bacteria degraded 63% of the total alkane and 66% the total PAHs. And the bacteria immobilized with the carriers degraded more HMW-alkanes and HMW-PAHs than the free bacteria. The bacteria immobilized by non-sterilized kelp residue showed a considerably higher degradation rate than that using sterilized kelp residue. A considerably higher cells absorption rate of immobilization was obtained when using kelp residue, and the preparation of immobilization was low cost and highly efficient. The experiments show the two algae materials, especially the kelp residue, present potential application in bioremediation of marine oil spills.

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Enzyme is an efficient and green biocatalyst, and widely used in many areas. Immobilized enzyme is superior to its free form in a variety of properties. Enzyme immobilization studies started in the 1970s in China. Till now, immobilized enzymes are widely applied in the fields of food, medical, energy, environmental management, among others. However, there are still some defects such as no universal method and the high cost. Therefore, based on the relatively mature traditional immobilization technologies, efforts have been made to innovate immobilization technologies. As a result, many new immobilization technologies focusing on new carriers and methods are continuously generated. Coupling with more than ten years' study on enzyme immobilization, we present here recent development and application of new immobilization technologies, as well as suggestions to future development of immobilization technology.

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Enzyme is an efficient and green biocatalyst, and widely used in many areas. Immobilized enzyme is superior to its free form in a variety of properties. Enzyme immobilization studies started in the 1970s in China. Till now, immobilized enzymes are widely applied in the fields of food, medical, energy, environmental management, among others. However, there are still some defects such as no universal method and the high cost. Therefore, based on the relatively mature traditional immobilization technologies, efforts have been made to innovate immobilization technologies. As a result, many new immobilization technologies focusing on new carriers and methods are continuously generated. Coupling with more than ten years' study on enzyme immobilization, we present here recent development and application of new immobilization technologies, as well as suggestions to future development of immobilization technology.