Regeneration System Research Articles

Lateral bud induction and establishment of regeneration system of cassia mimosoides L.

To establish an efficient plant regeneration system of Cassia mimosoides, the 10-day-old seedlings were used as explants for inducing lateral buds. Results showed that the addition of BA and NAA for different ratios on the basis of 1/2 MS could effectively induce the production of lateral buds. By adding different additional elements, it was found that the growth of lateral buds was longer and stronger on the medium of 1/2 MS + 3.0 mg/l BA + 0.1 mg/l NAA + 4.0 mg/l Gln or 1/2 MS + 3.0 mg/l BA + 0.1 mg/l NAA + 1.0 mg/l AgNO3. Meanwhile, cutting with the lateral buds of tissue culture seedling, the rooting rate could reach 77.8% when 1/2 MS medium contained 0.2 mg/l NAA, although the root length was shorter. When the regenerated strong seedlings were transplanted into the soil, the survival rate was over 75%. Therefore, the best medium for lateral bud induction using 10-day-old seedlings as explants in C. mimosoides was 1/2 MS + 3.0 mg/l BA + 0.1 mg/l NAA. The growth of lateral buds can be promoted by the appropriate concentration of Gln and AgNO3, and the best rooting medium was 1/2 MS contained 0.2 mg/l NAA. Bangladesh J. Bot. 52(2): 515-521, 2023 (June) Special

Lateral bud induction and establishment of regeneration system of cassia mimosoides L.

To establish an efficient plant regeneration system of Cassia mimosoides, the 10-day-old seedlings were used as explants for inducing lateral buds. Results showed that the addition of BA and NAA for different ratios on the basis of 1/2 MS could effectively induce the production of lateral buds. By adding different additional elements, it was found that the growth of lateral buds was longer and stronger on the medium of 1/2 MS + 3.0 mg/l BA + 0.1 mg/l NAA + 4.0 mg/l Gln or 1/2 MS + 3.0 mg/l BA + 0.1 mg/l NAA + 1.0 mg/l AgNO3. Meanwhile, cutting with the lateral buds of tissue culture seedling, the rooting rate could reach 77.8% when 1/2 MS medium contained 0.2 mg/l NAA, although the root length was shorter. When the regenerated strong seedlings were transplanted into the soil, the survival rate was over 75%. Therefore, the best medium for lateral bud induction using 10-day-old seedlings as explants in C. mimosoides was 1/2 MS + 3.0 mg/l BA + 0.1 mg/l NAA. The growth of lateral buds can be promoted by the appropriate concentration of Gln and AgNO3, and the best rooting medium was 1/2 MS contained 0.2 mg/l NAA. Bangladesh J. Bot. 52(2): 515-521, 2023 (June) Special

Structure-activity relationship of 2,4-D correlates auxinic activity with the induction of somatic embryogenesis in Arabidopsis thaliana.

2,4-dichlorophenoxyacetic acid (2,4-D) is a synthetic analogue of the plant hormone auxin that is commonly used in many in vitro plant regeneration systems, such as somatic embryogenesis (SE). Its effectiveness in inducing SE, compared to the natural auxin indole-3-acetic acid (IAA), has been attributed to the stress triggered by this compound rather than its auxinic activity. However, this hypothesis has never been thoroughly tested. Here we used a library of forty 2,4-D analogues to test the structure-activity relationship with respect to the capacity to induce SE and auxinic activity in Arabidopsis thaliana. Four analogues induced SE as effectively as 2,4-D and 13 analogues induced SE but were less effective. Based on root growth inhibition and auxin response reporter expression, the 2,4-D analogues were classified into different groups, ranging from very active to not active auxin analogues. A halogen at the 4-position of the aromatic ring was important for auxinic activity, whereas a halogen at the 3-position resulted in reduced activity. Moreover, a small substitution at the carboxylate chain was tolerated, as was extending the carboxylate chain with an even number of carbons. The auxinic activity of most 2,4-D analogues was consistent with their simulated TIR1-Aux/IAA coreceptor binding characteristics. A strong correlation was observed between SE induction efficiency and auxinic activity, which is in line with our observation that 2,4-D-induced SE and stress both require TIR1/AFB auxin co-receptor function. Our data indicate that the stress-related effects triggered by 2,4-D and considered important for SE induction are downstream of auxin signalling.

Innovative powertrain and advanced energy management strategy for hybrid hydraulic excavators

This paper presents an innovative powertrain managed by an advanced energy management strategy for hybrid hydraulic excavators to improve the overall energy efficiency. Here, a main pump is driven via a planetary gear by an engine and an electric motor/generator. A double clutch is integrated to adjust torque and speed of engine according to efficiency map enabling higher working efficiency. Potential energy of the boom cylinder can be converted and stored in a battery through an energy regeneration system. The advanced energy management strategy is designed by utilising extremum seeking and fuzzy techniques to optimally distribute power requirement. A fuzzy logic system is designed based on consideration of battery performance and aging to adjust a penalty function subsequently improving the overall system performance while prolonging the component lifetime. Simulations and real-time experiments have been carried out to evaluate the effectiveness of the proposed powertrain managed by the advanced energy management strategy.

Engineering a Formate Dehydrogenase for NADPH Regeneration.

Nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH) constitute major hydrogen donors for oxidative/reductive bio-transformations. NAD(P)H regeneration systems coupled with formate dehydrogenases (FDHs) represent a dreamful method. However, most of the native FDHs are NAD+ -dependent and suffer from insufficient reactivity compared to other enzymatic tools, such as glucose dehydrogenase. An efficient and competitive NADP+ -utilizing FDH necessitates the availability and robustness of NADPH regeneration systems. Herein, we report the engineering of a new FDH from Candida dubliniensis (CdFDH), which showed no strict NAD+ preference by a structure-guided rational/semi-rational design. A combinatorial mutant CdFDH-M4 (D197Q/Y198R/Q199N/A372S/K371T/▵Q375/K167R/H16L/K159R) exhibited 75-fold intensification of catalytic efficiency (kcat /Km ). Moreover, CdFDH-M4 has been successfully employed in diverse asymmetric oxidative/reductive processes with cofactor total turnover numbers (TTNs) ranging from 135 to 986, making it potentially useful for NADPH-required biocatalytic transformations.

High Voltage Long-Lifetime Mild pH-Decoupling Aqueous Flow Battery with in-Situ pH Recovery

Aqueous redox flow batteries (ARFBs) are being developed for grid scale electricity storage. However, the 1.23 V water splitting window makes the development of a long lifetime high-voltage ARFB challenging. Though the pH-decoupling design, with an acidic posolyte and an alkaline negolyte has been demonstrated to broaden the operating voltage window, long term operational stability and overall energy efficiency are penalized because of acid-base leakage along the pH gradient. Mild acid and base are used as supporting electrolytes to diminish proton and hydroxide crossover rates, resulting in higher overall energy efficiency. An acid-base in-situ regeneration system is implemented to restore the negolyte and posolyte pH to starting values. The capacity fade rate achieved is < 0.08%/day, with > 75% overall energy efficiency, and the Coulombic efficiency is around 99%. This work demonstrates a possibility of solving the critical issues (lifespan, rate capability, long term practicability and energy efficiency) for pH-decoupling ARFBs and guides the design for next generation high voltage ARFBs.

Rational design of polyphosphate kinase dual-substrate channel cavity for efficient production of glutathione by cell free catalysis

ATP is an important cofactor involved in many biocatalytic reactions that require energy input. Polyphosphate kinases (PPK) can provide energy for ATP-consuming reactions due to their cheap and readily available substrate polyphosphate. We selected ChPPK from Cytophaga hutchinsonii for substrate profiling and tolerance analysis. By molecular docking and site-directed mutagenesis, we rationally engineered the dual-substrate channel cavity of polyphosphate kinase to improve the catalytic activity of PPK. Compared with the wild type, the relative enzyme activity of the screened mutant ChPPKK81H-K103V increased by 326.7%. Meanwhile, the double mutation expanded the substrate utilization range and tolerance of ChPPK, and improved its heat and alkali resistance. Subsequently, we coupled the glutathione bifunctional enzyme GshAB and ChPPKK81H-K103V based on this ATP regeneration system, and glutathione was produced by cell-free catalysis upon disruption of cells. This system produced (25.4±1.9) mmol/L glutathione in 6 h upon addition of 5 mmol/L ATP. Compared with the system before mutation, glutathione production was increased by 41.9%. After optimizing the buffer, bacterial mass and feeding time of this system, (45.2±1.8) mmol/L glutathione was produced in 6 h and the conversion rate of the substrate l-cysteine was 90.4%. Increasing the ability of ChPPK enzyme to produce ATP can effectively enhance the conversion rate of substrate and reduce the catalytic cost, achieving high yield, high conversion rate and high economic value for glutathione production by cell-free catalysis. This study provides a green and efficient ATP regeneration system that may further power the ATP-consuming biocatalytic reaction platform.

Efficient photocatalytic NADH regeneration with Rh-loaded Z-scheme mediator-free system

Nicotinamide adenine dinucleotide (NADH) regeneration is necessary for the sustainable application of enzymatic industry. The Rh-based complex [Cp*Rh(bpy)(H)]+ has been widely used as an important mediator in NADH regeneration systems, but it is limited by complexity and high cost. Here, a Z-scheme was constructed by loading Rh onto carbon nitride nanosheets/carbon nitride quantum dots (CN-CNQD). The resultant catalyst achieved a high yield of NADH in a mediator-free (M-free) system of 0.283 mmol L−1 g−1 min−1, which is 5.29 times that of pure CN. ADH enzyme introduction experiments confirmed that the enzyme active product 1,4-NADH could reach 34.21% selectivity in the M-free system. Mechanism research revealed that the heterojunction between CNs and CNQDs improved the NADH regeneration activity in the traditional M-involved system, while Rh loading was proved to optimize the yield and selectivity of 1,4-NADH in M-free system. The immobilized Rh shows more competitiveness than [Cp*Rh(bpy)(H)]+. This study contributes to the construction of an M-free system for further application in greener, lower-cost enzymatic processes.

Power output of magnetocaloric materials during heat and cold decoupling by unidirectional fluid flow

This article presents the results of a novel concept of heat and cold decoupling from magnetocaloric materials (MCM) by unidirectional fluid flow. An experimental setup was developed to analyse the heat/cold decoupling and determine the optimal process parameters. Three different kinds of magnetocaloric material samples were analysed by using this setup. Comparing the measuring results to those of existing active magnetic regenerator (AMR) systems shows a very good performance and indicates a higher achievable thermal power output when using a unidirectional fluid flow.

A novel salt- and organic solvent-tolerant phosphite dehydrogenase from Cyanothece sp. ATCC 51142.

Phosphite dehydrogenase (PtxD) is a promising enzyme for NAD(P)H regeneration. To expand the usability of PtxD, we cloned, expressed, and analyzed PtxD from the marine cyanobacterium Cyanothece sp. ATCC 51142 (Ct-PtxD). Ct-PtxD exhibited maximum activity at pH 9.0°C and 50°C and high stability over a wide pH range of 6.0-10.0. Compared to previously reported PtxDs, Ct-PtxD showed increased resistance to salt ions such as Na+, K+, and NH4 +. It also exhibited high tolerance to organic solvents such as ethanol, dimethylformamide, and methanol when bound to its preferred cofactor, NAD+. Remarkably, these organic solvents enhanced the Ct-PtxD activity while inhibiting the PtxD activity of Ralstonia sp. 4506 (Rs-PtxD) at concentrations ranging from 10% to 30%. Molecular electrostatic potential analysis showed that the NAD+-binding site of Ct-PtxD was rich in positively charged residues, which may attract the negatively charged pyrophosphate group of NAD+ under high-salt conditions. Amino acid composition analysis revealed that Ct-PtxD contained fewer hydrophobic amino acids than other PtxD enzymes, which reduced the hydrophobicity and increased the hydration of protein surface under low water activity. We also demonstrated that the NADH regeneration system using Ct-PtxD is useful for the coupled chiral conversion of trimethylpyruvic acid into L-tert-leucine using leucine dehydrogenase under high ammonium conditions, which is less supported by the Rs-PtxD enzyme. These results imply that Ct-PtxD might be a potential candidate for NAD(P)H regeneration in industrial applications under the reaction conditions containing salt and organic solvent.

Influence of Thidiazuron on in vitro regeneration potential of Kaempferia parviflora Wall. Ex Baker from Eastern India

An in vitro regeneration system was standarized on a traditional medicinal plant of Kaempferia parviflora. The rhizome sprouted buds upon transfer to MS basal medium with kinetin (3mg/l) and naphthalene acetic acid (0.5mg/l) were found optimum exhibiting maximum percent of shoot initiation as 90.6 ± 0.2. Maximum number of shoots as 8.1 ± 0.3 and maximum number of roots as 22.0 ± 0.3 were found in benzyladenine (3mg/l), naphthalene acetic acid (1mg/l) and thidiazuron (0.5mg/l) combination. Then the micropropagated plants were transferred to field for successful establishment as 90%. Furthermore, the morphological characteristic analysis was done between the conventional and micropropagated plants which showed very less significant variation. Thus the present protocol reports the propagation of K. parviflora plantlets which could be used for commercial exploitation in future.

Role of the Peripheral Nervous System in Skeletal Development and Regeneration: Controversies and Clinical Implications.

This review examines the diverse functional relationships that exist between the peripheral nervous system (PNS) and bone, including key advances over the past century that inform our efforts to translate these discoveries for skeletal repair. The innervation of the bone during development, homeostasis, and regeneration is highly patterned. Consistent with this, there have been nearly 100 studies over the past century that have used denervation approaches to isolate the effects of the different branches of the PNS on the bone. Overall, a common theme of balance emerges whereby an orchestration of both local and systemic neural functions must align to promote optimal skeletal repair while limiting negative consequences such as pain. An improved understanding of the functional bidirectional pathways linking the PNS and bone has important implications for skeletal development and regeneration. Clinical advances over the next century will necessitate a rigorous identification of the mechanisms underlying these effects that is cautious not to oversimplify the in vivo condition in diverse states of health and disease.

Complementation of reducing power for 5-hydroxyvaleric acid and 1,5-pentanediol production via glucose dehydrogenase in Escherichia coli whole-cell system

One of the key intermediates, 5-hydroxyvaleric acid (5-HV), is used in the synthesis of polyhydroxyalkanoate monomer, δ-valerolactone, 1,5-pentanediol (1,5-PDO), and many other substances. Due to global environmental problems, eco-friendly bio-based synthesis of various platform chemicals and key intermediates are socially required, but few previous studies on 5-HV biosynthesis have been conducted. To establish a sustainable bioprocess for 5-HV production, we introduced gabT encoding 4-aminobutyrate aminotransferase and yqhD encoding alcohol dehydrogenase to produce 5-HV from 5-aminovaleric acid (5-AVA), through glutarate semialdehyde in Escherichia coli whole-cell reaction. As, high reducing power is required to produce high concentrations of 5-HV, we newly introduced glucose dehydrogenase (GDH) for NADPH regeneration system from Bacillus subtilis 168. By applying GDH with D-glucose and optimizing the parameters, 5-HV conversion rate from 5-AVA increased from 47% (w/o GDH) to 82% when using 200 mM (23.4 g/L) of 5-AVA. Also, it reached 56% conversion in 2 h, showing 56 mM/h (6.547 g/L/h) productivity from 200 mM 5-AVA, finally reaching 350 mM (41 g/L) and 14.6 mM/h (1.708 g/L/h) productivity at 24 h when 1 M (117.15 g/L) 5-AVA was used. When the whole-cell system with GDH was expanded to produce 1,5-PDO, its production was also increased 5-fold. Considering that 5-HV and 1,5-PDO production depends heavily on the reducing power of the cells, we successfully achieved a significant increase in 5-HV and 1,5-PDO production using GDH.

The salamander blastema within the broader context of metazoan regeneration.

Throughout the animal kingdom regenerative ability varies greatly from species to species, and even tissue to tissue within the same organism. The sheer diversity of structures and mechanisms renders a thorough comparison of molecular processes truly daunting. Are "blastemas" found in organisms as distantly related as planarians and axolotls derived from the same ancestral process, or did they arise convergently and independently? Is a mouse digit tip blastema orthologous to a salamander limb blastema? In other fields, the thorough characterization of a reference model has greatly facilitated these comparisons. For example, the amphibian Spemann-Mangold organizer has served as an amazingly useful comparative template within the field of developmental biology, allowing researchers to draw analogies between distantly related species, and developmental processes which are superficially quite different. The salamander limb blastema may serve as the best starting point for a comparative analysis of regeneration, as it has been characterized by over 200years of research and is supported by a growing arsenal of molecular tools. The anatomical and evolutionary closeness of the salamander and human limb also add value from a translational and therapeutic standpoint. Tracing the evolutionary origins of the salamander blastema, and its relatedness to other regenerative processes throughout the animal kingdom, will both enhance our basic biological understanding of regeneration and inform our selection of regenerative model systems.

Systematic Engineering of Saccharomyces cerevisiae Chassis for Efficient Flavonoid-7-O-Disaccharide Biosynthesis.

Flavonoids are an essential class of secondary metabolites found in plants and possess various nutritional, medicinal, and agricultural properties. However, the poor water solubility of flavonoid aglycones limits their potential applications. To overcome this issue, glycosylation is a promising approach for improving water solubility and bioavailability. In this study, we constructed a flavonoid-7-O-disaccharide biosynthetic pathway with flavonoid aglycones as substrates in Saccharomyces cerevisiae. Subsequently, through metabolic engineering and promoter strategies, we constructed a UDP-rhamnose regeneration system and optimized the UDP-glucose (UDPG) synthetic pathway. The optimized strain produced up to 131.3 mg/L eriocitrin. After this, the chassis cells were applied to other flavonoids, with substrates such as (2S)-naringenin, (2S)-hesperetin, diosmetin, and (2S)-eriodictyol, which resulted in the synthesis of 179.9 mg/L naringin, 276.6 mg/L hesperidin, 249.0 mg/L neohesperidin, 30.4 mg/L diosmin, and 100.7 mg/L neoeriocitrin. To the best of our knowledge, this is the first report on the biosynthesis of flavonoid-7-O-disaccharide.

Enhancing drought resistance in Pogostemon cablin (Blanco) Benth. through overexpression of ACC deaminase gene using thin cell layer regeneration system.

Pogostemon cablin cultivation faces massive constraints because of its susceptability to drought stress that reduces patchouli propagation and oil yield. The present study has achieved an efficient and rapid direct regeneration system for the transgenic production of P. cablin using Agrobacterium-mediated genetic transformation. To establish an efficient regeneration protocol for fast in-vitro multiplication of patchouli plants, leaf, petiole, and transverse thin cell layer (tTCL) explants were used and inoculated on an MS medium supplemented with different combinations of phytohormones. A comparative study showed a maximum regeneration frequency of 93.30 ± 0.56% per explant was obtained from leaf segments on optimal MS medium fortified with 0.2mg/L BAP and 0.1mg/L NAA. Leaf and petiole explants took 25-35 days to regenerate while tTCL section showed regeneration in just 15-20 days on the same medium. Subsequently, productive genetic transformation protocol OD600 0.6, AS 200µM, 30mg/L kanamycin, and infection time 5 min. was standardized and best-suited explants were infected at optimum conditions from the Agrobacterium tumefaciens (LBA 4404) strain harboring ACC deaminase to generate transgenic P. cablin Benth. (CIM-Samarth) plants. The investigation suggested that the optimized protocol provides a maximum transformation frequency of 42 ± 1.9% in 15-20 days from tTCL. The transgenic plants were shifted to the greenhouse with a 52.0 ± 0.8% survival frequency. A molecular docking study confirmed significant binding affinity of ligand ACC with ACC deaminase at the catalytic site, and ligand interactions showed four H-bonds at the binding pocket with amino acids Cys-196, Val-198, Thr-199, and Gly-200 that validate gene relative expression in transgenic plants. Among all transgenic acclimatized greenhouse-grown patchouli plants, line PT4 showed improved drought resistance under severe water stress as its RWC was 71.7 ± 2.3% to 75.7 ± 2.1% which is greater than the RWC of the control plant, 58.30 ± 0.21%. Analysis of the other physiological indicators, H2O2, chlorophyll content, and ROS result support drought resistance ability. Our study concluded that the first report on P. cablin, tTCL direct regeneration, and standardized transformation protocol created a new opportunity for genetic manipulation to achieve drought-resistant patchouli plants for cultivation in all seasons at the commercial level.

An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis.

The Agrobacterium tumefaciens-mediated transformation for blueberries remains less efficient than is desirable. A new leaf callus regeneration and genetic transformation system was investigated in blueberries in this study. The leaf explants of cv. 'Legacy' and 'Northland' were used to establish the stable callus induction system when placed on the woody plant medium (WPM) supplemented with 1.0 mg·L-1 2, 4-D, 0.4 mg·L-1 6-BA for 30 d; then, the callus was sub-cultured in the proliferation medium supplemented with 1.5 mg·L-1 2, 4-D, 0.4 mg·L-1 6-BA in the darkness at 25 °C every 30 days. The co-cultivation of callus with A. tumefaciens was operated on WPM plus 100 μM acetosyringone for 4 days; then, the transferred callus was grown in WPM supplemented with 1.5 mg·L-1 2,4-D, 0.4 mg·L-1 6-BA, 50 mg·L-1 hygromycin, and 200 mg·L-1 cefotaxime. The VcCHS transgenic blueberry callus with both GFP signal and Hyg resistance was obtained from the transformed callus of cv. 'Northland'. The rate of GFP signal detected in the transformed callus was as high as 49.02%, which was consistent with the PCR assay. Collectively, this study provides a highly efficient genetic transformation system in blueberry callus and a powerful approach for the molecular breeding of blueberries.

Fabrication of avian eggshell membrane derived dispersed collagen hydrogels for potential bone regeneration

Abstract Tissue engineering is emerging as an effective and alternate strategy for healing the impaired bones. This study reports the development of potential and novel bone tissue regenerating matrices from the avian eggshell membrane derived collagen dispersed in poly-(vinyl alcohol) (PVA). The concentration of the aqueous collagen dispersions within the PVA solution were varied from 0.5 % to 2 % (w/V); and the sols were subjected to varying freeze-thaw cycles to achieve gelation; leading to the fabrication of hydrogel matrices. The developed hydrogels were investigated for their physico-chemical characteristics through the ATR-FTIR, XRD, FESEM and biocompatibility measurements. The ATR-FTIR results showed the presence of amide A, amide I and hydroxyl groups; which were further reinstated by the XRD studies. The morphological and topological analysis of the different hydrogel groups was made through FESEM. It comprised of the measurements of pore dimensions ranging (3.98 ± 1.84 µm to 9.24 ± 5.55 µm), percentage porosity (47 %–97 %), and average surface roughness (21–39 µm); thus, indicating them to be analogous support systems for bone tissue regeneration. Further, L929 mouse fibroblasts grown over these support systems showed excellent cell viability, thus hinting towards its competitive features and application towards remedial bone regeneration.

Continuous cultivation of microalgae yields high nutrient recovery from nitrified urine with limited supplementation

Microalgae can play a key role in the bioeconomy, particularly in combination with the valorisation of waste streams as cultivation media. Urine is an example of a widely available nutrient-rich waste stream, and alkaline stabilization and subsequent full nitrification in a bioreactor yields a stable nitrate-rich solution. In this study, such nitrified urine served as a culture medium for the edible microalga Limnospira indica. In batch cultivation, nitrified urine without additional supplements yielded a lower biomass concentration, nutrient uptake and protein content compared to modified Zarrouk medium, as standard medium. To enhance the nitrogen uptake efficiency and biomass production, nitrified urine was supplemented with potentially limiting elements. Limited amounts of phosphorus (36 mg L−1), magnesium (7.9 mg L−1), calcium (12.2 mg L−1), iron (2.0 mg L−1) and EDTA (88.5 mg Na2-EDTA.2H2O L−1) rendered the nitrified urine matrix as effective as modified Zarrouk medium in terms of biomass production (OD750 of 1.2), nutrient uptake (130 mg N L−1) and protein yield (47%) in batch culture. Urine precipitates formed by alkalinisation could in principle supply enough phosphorus, calcium and magnesium, requiring only external addition of iron, EDTA and inorganic carbon. Subsequently, the suitability of supplemented nitrified urine as a culture medium was confirmed in continuous Limnospira cultivation in a CSTR photobioreactor. This qualifies nitrified urine as a valuable and sustainable microalgae growth medium, thereby creating novel nutrient loops on Earth and in Space, i.e., in regenerative life support systems for human deep-space missions.

A high-throughput dual system to screen polyphosphate kinase mutants for efficient ATP regeneration in L-theanine biocatalysis

ATP, an important cofactor, is involved in many biocatalytic reactions that require energy. Polyphosphate kinases (PPK) can provide energy for ATP-consuming reactions due to their cheap and readily available substrate polyphosphate. We determined the catalytic properties of PPK from different sources and found that PPK from Cytophaga hutchinsonii (ChPPK) had the best catalytic activity for the substrates ADP and polyP6. An extracellular–intracellular dual system was constructed to high-throughput screen for better catalytic activity of ChPPK mutants. Finally, the specific activity of ChPPKD82N-K103E mutant was increased by 4.3 times. Therefore, we focused on the production of L-theanine catalyzed by GMAS as a model of ATP regeneration. Supplying 150 mM ATP, GMAS enzyme could produce 16.8 ± 1.3 g/L L-theanine from 100 mM glutamate. When 5 mM ATP and 5 U/mL ChPPKD82N-K103E were added, the yield of L-theanine was 16.6 ± 0.79 g/L with the conversion rate of 95.6 ± 4.5% at 4 h. Subsequently, this system was scaled up to 200 mM and 400 mM glutamate, resulting in the yields of L-theanine for 32.3 ± 1.6 g/L and 62.7 ± 1.1 g/L, with the conversion rate of 92.8 ± 4.6% and 90.1 ± 1.6%, respectively. In addition, we also constructed an efficient ATP regeneration system from glutamate to glutamine, and 13.8 ± 0.2 g/L glutamine was obtained with the conversion rate of 94.4 ± 1.4% in 4 h after adding 6 U/ mL GS enzyme and 5 U/ mL ChPPKD82N-K103E, which further laid the foundation from glutamine to L-theanine catalyzed by GGT enzyme. This proved that giving the reaction an efficient ATP supply driven by the mutant enzyme enhanced the conversion rate of substrate to product and maximized the substrate value. This is a positively combination of high yield, high conversion rate and high economic value of enzyme catalysis. The mutant enzyme will further power the ATP-consuming biocatalytic reaction platform sustainably.