Detection and elimination of trace d-lactic acid in lignocellulose biorefining chain: Generation, flow, and impact on chiral lactide synthesis.

High chiral purity of lactic acid is a crucial indicator for the synthesis of chiral lactide as the primary intermediate chemical for ring-open polymerization of high molecular weight polylactic acid (PLA). Lignocellulose biomass is the most promising carbohydrate feedstock for commercial production of PLA, but the presence of trace d-lactic acid in the biorefinery chain adversely affects the synthesis and quality of chiral lactide. This study analyzed the fingerprint of trace  d-lactic acid in the biorefinery chain and found that the major source of  d-lactic acid comes from lignocellulose feedstock. The naturally occurring lactic acid bacteria and water-soluble carbohydrates in lignocellulose feedstock provide the necessary conditions for  d-lactic acid generation. Three strategies were proposed to eliminate the generation pathway of  d-lactic acid, including reduction of moisture content, conversion of water-soluble carbohydrates to furan aldehydes in pretreatment, and conversion to  l-lactic acid by inoculating engineered  l-lactic acid bacteria. The natural reduction of lactic acid content in lignocellulose feedstock during storage was observed due to the lactate oxidase-catalyzed oxidation of  l- and  d-lactic acids. This study provided an important support for the production of cellulosic  l-lactic acid with high chiral purity.

Highly efficient and recyclable monolithic bioreactor for interfacial enzyme catalysis.

HYPOTHESIS: Biocatalysts are key to the realization of all bioconversions in nature. However, the difficulty of combining the biocatalyst and other chemicals in one system limits their application in artificial reaction systems. Although some effort, such as Pickering interfacial catalysis and enzyme-immobilized microchannel reactors, have addressed this challenge an effective method to combine chemical substrates and biocatalysts in a highly efficient and re-usable monolith system is still to be developed. EXPERIMENTS: A repeated batch-type biphasic interfacial biocatalysis microreactor was developed using enzyme-loaded polymersomes in the void surface of porous monoliths. Polymersomes, loaded with Candida antarctica Lipase B (CALB), are fabricated by self-assembly of the copolymer PEO-b-P(St-co-TMI) and used to stabilize oil-in-water (o/w) Pickering emulsions as a template to prepare monoliths. By adding monomer and Tween 85 to the continuous phase, controllable open-cell monoliths are prepared to inlay CALB-loaded polymersomes in the pore walls. FINDINGS: The microreactor is proven to be highly effective and recyclable when a substrate flows through it, which offers superior benefits of absolute separation to a pure product and no enzyme loss. The relative enzyme activity is constantly maintained above 93% in 15 cycles. The enzyme is constantly present in the microenvironment of the PBS buffer ensuring its immunity to inactivation and facilitating its recycling.

Coproduction of single cell protein and lipid from lignocellulose derived carbohydrates and inorganic ammonia salt with soluble ammonia recycling.

Co-production of single cell protein (SCP) and lipid from lignocellulose-derived carbohydrates and inorganic ammonia offers a promising alternative for poultry or aquaculture feeds. An engineered oleaginous yeast Trichosporon cutaneum MP11 showed great potential for producing SCP and lipid from wheat straw and ammonia sulfate with minimum nutrient input. Trichosporon cutaneum MP11 showed stronger SCP and lipid fermentability using dry acid pretreated and biodetoxified wheat straw than using pure sugars. The residual ammonium sulfate in fermentation broth was recycled up to five times, resulting in ∼70% of nitrogen fixation into SCP. The overall yield of SCP and lipid from lignocellulose-derived sugars was 0.15 g/g and 0.11 g/g, respectively. This translates to the production of one ton of SCP (0.56 ton) and lipid (0.44 ton) from 6.6 tons of wheat straw, or one ton of SCP and lipid containing yeast cells (dry) from 4.8 tons of wheat straw.

Hierarchical Emulsion-Templated Monoliths (polyHIPEs) as Scaffolds for Covalent Immobilization of P. acidilactici.

The immobilized cell fermentation technique (IMCF) has gained immense popularity in recent years due to its capacity to enhance metabolic efficiency, cell stability, and product separation during fermentation. Porous carriers used as cell immobilization facilitate mass transfer and isolate the cells from an adverse external environment, thus accelerating cell growth and metabolism. However, creating a cell-immobilized porous carrier that guarantees both mechanical strength and cell stability remains challenging. Herein, templated by water-in-oil (w/o) high internal phase emulsions (HIPE), we established a tunable open-cell polymeric P(St-co-GMA) monolith as a scaffold for the efficient immobilization of Pediococcus acidilactici (P. acidilactici). The porous framework's mechanical property was substantially improved by incorporating the styrene monomer and cross-linker divinylbenzene (DVB) in the HIPE's external phase, while the epoxy groups on glycidyl methacrylate (GMA) supply anchoring sites for P. acidilactici, securing the immobilization to the inner wall surface of the void. For the fermentation of immobilized P. acidilactici, the polyHIPEs permit efficient mass transfer, which increases along with increased interconnectivity of the monolith, resulting in higher L-lactic acid yield compared to that of suspended cells with an increase of 17%. The relative L-lactic acid production is constantly maintained above 92.9% of their initial relative production after 10 cycles, exhibiting both its great cycling stability and the durability of the material structure. Furthermore, the procedure during recycle batch also simplifies downstream separation operations.

Simultaneous and rate-coordinated conversion of lignocellulose derived glucose, xylose, arabinose, mannose, and galactose into D-lactic acid production facilitates D-lactide synthesis.

D-lactide is the precursor of poly(D-lactide) (PDLA) or stereo-complex with poly(L-lactide) (PLLA). Lignocellulosic biomass provides the essential feedstock option to synthesize D-lactic acid and D-lactide. The residual sugars in D-lactic acid fermentation broth significantly blocks the D-lactide synthesis. This study showed a simultaneous and rate-coordinated conversion of lignocellulose derived glucose, xylose, arabinose, mannose, and galactose into D-lactic acid by adaptively evolved Pediococcus acidilactici ZY271 by simultaneous saccharification and co-fermentation (SSCF) of wheat straw. The produced D-lactic acid achieved minimum residual sugars (∼1.7 g/L), high chirality (∼99.1%) and high titer (∼128 g/L). A dry acid pretreatment eliminated the wastewater stream generation and the biodetoxification by fungus Amorphotheca resinae ZN1 removed the inhibitors from the pretreatment. The removal of the sugar residues and inhibitor impurities in D-lactic acid production from lignocellulose strongly facilitated the D-lactide synthesis. This study filled the gap in cellulosic D-lactide production from lignocellulose-derived D-lactic acid.

High solids loading pretreatment: The core of lignocellulose biorefinery as an industrial technology - An overview.

Pretreatment is the first and most determinative, yet the least mature step of lignocellulose biorefinery chain. The current stagnation of biorefinery commercialization indicates the barriers of the existing pretreatment technologies are needed to be unlocked. This review focused on one of the core factors, the high lignocellulose solids loading in pretreatment. The high solids loading of pretreatment significantly reduces water input, energy requirement, toxic compound discharge, solid/liquid separation costs, and carbon dioxide emissions, improves the titers of sugars and biproducts to meet the industrial requirements. Meanwhile, lignocellulose feedstock after high solids loading pretreatment is compatible with the existing logistics system for densification, packaging, storage, and transportation. Both the technical-economic analysis and the cellulosic ethanol conversion performance suggest that the solids loading in the pretreatment step need to be further elevated towards an industrial technology and the effective solutions should be proposed to the technical barriers in high solids loading pretreatment operations.

Effect of Antioxidants on Thermo-Oxidative Stability and Aging of Bio-Based PA56T and Fast Characterization of Anti-Oxidation Performance.

Bio-based polyamide 56T (PA56T) is a new type of bio-based polyamide regarded as a promising material for sustainable solutions. The stabilization of PA56T compounded with Irganox 1098, Doverphos S9228, or SH3368 was studied by using a rotational rheometer and a circulating air oven at 150 °C. The thermal-oxidative aging resulted in an increase of the yellow color index of the PA56T/GF composites, which due to the carbonyl group as a chromophore group, continuously formatted during the aging process. After 10 days of aging, the mechanical properties and dynamic mechanical properties increase due to the molecular cross-linking and annealing effects. When the aging time is beyond 20 days, the degradation of molecular chain segments dominates, and the mechanical properties of PA56T/GF deteriorate continuously. The addition of antioxidants only slowed this effect and did not change the process of thermal-oxidative aging, which destroys the molecular chain. The results from both methods are consistent after a series of characterizations by FTIR, XRD, and so on. In the case of samples without lubricant, the rotational rheometer has the benefit of being less time-consuming than the accelerated aging experiment.

Cyclic l-lactide synthesis from lignocellulose biomass by biorefining with complete inhibitor removal and highly simultaneous sugars assimilation.

Cyclic chiral lactide is the monomer chemical for polymerization of high molecular weight polylactic acid (PLA). The synthesis of cyclic l-lactide starts from poly-condensation of l-lactic acid to a low molecular weight prepolymer and then depolymerized to cyclic l-lactide. Lignocellulose biomass is the most promising carbohydrate feedstock for lactic acid production, but the synthesis of cyclic l-lactide from l-lactic acid produced from lignocellulose has so far not been successful. The major barriers are the impurities of residual sugars and inhibitors in the crude cellulosic l-lactic acid product. Here we show a successful cyclic l-lactide synthesis from cellulosic l-lactic acid by lignocellulose biorefining with complete inhibitor removal and coordinated sugars assimilation. The removal of inhibitors from lignocellulose pretreatment was accomplished by biodetoxification using a unique fungus Amorphotheca resinae ZN1. The nonglucose sugars were completely and simultaneously assimilated at the same rate with glucose by the engineered l-lactic acid bacterium Pediococcus acidilactici. The l-lactic acid production from wheat straw was comparable to that from corn starch with high optical pure (99.6%), high l-lactic acid titer (129.4 g/L), minor residual total sugars (~2.2 g/L), and inhibitors free. The cyclic l-lactide was successfully synthesized from the regularly purified l-lactic acid and verified by detailed characterizations. This study paves the technical foundation of carbon-neutral production of biodegradable PLA from lignocellulose biomass.

Auto-classification of biomass through characterization of their pyrolysis behaviors using thermogravimetric analysis with support vector machine algorithm: case study for tobacco.

BACKGROUND: During the biomass-to-bio-oil conversion process, many studies focus on studying the association between biomass and bio-products using near-infrared spectra (NIR) and chemical analysis methods. However, the characterization of biomass pyrolysis behaviors using thermogravimetric analysis (TGA) with support vector machine (SVM) algorithm has not been reported. In this study, tobacco was chosen as the object for biomass, because the cigarette smoke (including water, tar, and gases) released by tobacco pyrolysis reactions decides the sensory quality, which is similar to biomass as a renewable resource through the pyrolysis process. RESULTS: SVM algorithm has been employed to automatically classify the planting area and growing position of tobacco leaves using thermogravimetric analysis data as the information source for the first time. Eighty-eight single-grade tobacco samples belonging to four grades and eight categories were split into the training, validation, and blind testing sets. Our model showed excellent performances in both the training and validation set as well as in the blind test, with accuracy over 91.67%. Throughout the whole dataset of 88 samples, our model not only provides precise results on the planting area of tobacco leave, but also accurately distinguishes the major grades among the upper, lower, and middle positions. The error only occurs in the classification of subgrades of the middle position. CONCLUSIONS: From the case study of tobacco, our results validated the feasibility of using TGA with SVM algorithm as an objective and fast method for auto-classification of tobacco planting area and growing position. In view of the high similarity between tobacco and other biomasses in the compositions and pyrolysis behaviors, this new protocol, which couples the TGA data with SVM algorithm, can potentially be extrapolated to the auto-classification of other biomass types.

Serum- and glucocorticoid-regulated protein kinase 3 overexpression promotes tumor development and aggression in breast cancer cells.

Serum- and glucocorticoid-regulated protein kinase 3 (SGK3) is critical for tumor survival, proliferation and invasion. In the present study we evaluated SGK3 expression in breast tissues and investigated alterations in SGK3 levels in tumor multiplication, progression and apoptosis. Tissue microarray analyses were performed to examine SGK3 expression in breast cancer samples, as well as in adjacent noncancerous and normal tissues. The pEGFP-N1-SGK3 plasmid was transfected into MDA-MB-231 cells to generate SGK3-overexpressing cells. Cell growth assays, colony formation assays, cell cycle analyses, horizontal and vertical migration tests, reverse transcription-polymerase chain reaction and western blot assays were employed to investigate the biological behavior of SGK3-overexpressing cells. SGK3 levels were significantly higher in breast cancer samples compared with adjacent noncancerous and normal tissues. Cell growth curves revealed increased proliferation and decreased apoptosis in SGK3-overexpressing cells. SGK3-overexpressing cells also demonstrated enhanced invasion and migration abilities. SGK3-overexpressing cells had high levels of an apoptosis-related gene (bcl-xl) and invasion-related genes (mmp2 and mmp9), and decreased levels of an anti-apoptosis gene (bad). Phosphorylation of GSK-3ß, which is downstream in the phosphoinositide 3-kinase signaling pathway, was activated by SGK3 overexpression. ß-catenin phosphorylation did not differ between the SGK3-overexpressing and non-SGK3-overexpressing cells. SGK3 overexpression induces GSK-3ß phosphorylation, enhancing apoptosis- and invasion-related genes and proteins and thereby leading to tumor development and aggression in breast cancer cells.

Protective effect and mechanism of glutaredoxin 1 on coronary arteries endothelial cells damage induced by high glucose.

In recent years, diabetes and its associated complications have become a major public health concern. The cardiovascular risk increases significantly in diabetes patients. It is a complex disease characterized by multiple metabolic derangements and is known to impair cardiac function by disrupting the balance between pro-oxidants and antioxidants at the cellular level. The subsequent generation of reactive oxygen species (ROS) and accompanying oxidative stress are hallmarks of the molecular mechanisms responsible for cardiovascular disease. Protein thiols act as redox-sensitive switches and are believed to be a key element in maintaining the cellular redox balance. The redox state of protein thiols is regulated by oxidative stress and redox signaling and is important to cellular functions. The potential of the thiol-disulfide oxidoreductase enzymes (thioredoxin and glutaredoxin systems) in defense against oxidative stress has been noted previously. Increasing evidence demonstrates that glutaredoxin 1 (Grx1), a cytosolic enzyme responsible for the catalysis of protein deglutathionylation, plays distinct roles in inflammation and apoptosis by inducing changes in the cellular redox system. This study investigates whether and how Grx1 protects coronary artery vascular endothelial cells against high glucose (HG) induced damage. Results indicate that the activation of eNOS/NO system is regulated by Grx 1 and coupled with inhibition of JNK and NF-κB signaling pathway which could alleviate the oxidative stress and apoptosis damage in coronary arteries endothelial cells induced by HG.