d -Tagatose production by permeabilized and immobilized Lactobacillus plantarum using whey permeate

Jayamuthunagai, J.; Srisowmeya, G.; Chakravarthy, M.; Pennathur, Gautam

doi:10.1016/j.biortech.2017.03.123

Cited by 49 publications

(28 citation statements)

References 27 publications

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“…Seeking to overcome the transport-limited whole-cell production of tagatose, we investigated the use of permeabilization surfactants − Triton X-100 and SDS. Following the same optimized protocol as previously reported for permeabilization of L. plantarum with 1 % Triton X-100 27 , we observed a 4.2 ± 0.1 fold increase in tagatose production as compared to untreated cells under the tested conditions (Fig. S4a).…”

Section: Resultssupporting

confidence: 59%

“…Cells were resuspended to n OD 600 = 3.0 in 200 μL of PBS. Samples were either incubated with 1% Triton X-100 27 , 0.1 % SDS, or no treatment at room temperature for 1 h. The solution was removed, and the cell pellet washed twice with PBS before enzymatic activity of the whole-cells were measured as described previously using 200 mM of glucose, galactose, or arabinose as substrates. Activity was quantified via the CCSAA and normalized to cell concentration as described previously.…”

Section: Methodsmentioning

confidence: 99%

“…Whole-cell biocatalysts with GRAS organisms (e.g. lactic acid bacteria (LAB) and E. coli ) that disproportionately partition substrate and product across their membrane has also been shown to partially circumvent this thermodynamic limitation while simultaneously enhancing enzyme stability; albeit at a kinetic penalty imposed by substrate transport limitations 25–27 . Recently, cell permeabilization 27 and sugar transport overexpression 28,29 were demonstrated as methods to overcome the kinetic penalty imposed by cellular encapsulation.…”

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confidence: 99%

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Surpassing thermodynamic, kinetic, and stability barriers to isomerization catalysis for tagatose biosynthesis

Bober

Nair

2019

Preprint

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3There are many enzymes that are relevant for making rare and valuable chemicals that 4 while active, are severely limited by thermodynamic, kinetic, or stability issues (e.g. 5 isomerases, lyases, transglycosidase etc.). In this work, we study an enzymatic reaction 6 system − Lactobacillus sakei L-arabinose isomerase (LsLAI) for D-galactose to D-tagatose 7 isomerizationthat is limited by all three reaction parameters. The enzyme has a low 8 catalytic efficiency for non-natural substrate galactose, has low thermal stability at 9 temperatures > 40 °C, and equilibrium conversion < 50%. After exploring several strategies 10 to overcome these limitations, we finally show that encapsulating the enzyme in a gram-11 positive bacterium (Lactobacillus plantarum) that is chemically permeabilized can enable 12 reactions at high rates, high conversion, and at high temperatures. The modified whole cell 13 system stabilizes the enzyme, differentially partitions substrate and product across the 14 membrane to shift the equilibrium toward product formation enables rapid transport of 15 substrate and product for fast kinetics. In a batch process, this system enables 16 approximately 50 % conversion in 4 h starting with 300 mM galactose (an average 17 productivity of 37 mM/h), and 85 % conversion in 48 h, which are the highest reported for 18 food-safe mesophilic tagatose synthesis. We suggest that such an approach may be 19 invaluable for other enzymatic processes that are similarly kinetically-, thermodynamically-20 , and/or stability-limited. 65We use the food-safe engineered probiotic bacterium Lactobacillus plantarum as the 66 expression host due to its increasing relevance to biochemical and biomedical research 30,31 . 67This approach enabled ~ 50 % conversion of galactose to tagatose in 4 h (productivity of ~ 68 38 mmol tagatose L -1 h -1 ) ultimately reaching ~ 85% conversion after 48 h at high galactose 69 loading (300 mM) in batch culture. This is among the highest conversions and 70 productivities reported to date for tagatose production using a mesophilic enzyme. Such an 71 approach is expected to be applicable to other biocatalytic systems where similar trade-offs 72 between kinetics, thermodynamics, and/or stability pose hurdles to process development. 73 74 Results: 75Characterizing L. sakei LAI (LsLAI) limitations. 76 At high substrate loading (400mM galactose) and 37 °C, the reported optimal temperature 77 for this enzyme 32 , LsLAI purified from L. plantarum exhibited an initial forward reaction 78 (turnover) rate (kinitial) of 9.3 ± 0.3 s -1 , which is lower than the maximum reaction rate 79 possible by this enzyme of 17.0 ± 1.3 s -1 min -1 with its preferred substrate, arabinose, at 400 80 mM. Increasing the reaction temperature to 50 °C increased the initial reaction rate to 11.2 81 s -1 ( Fig. 1a) but was accompanied by rapid enzyme inactivation, which is consistent with 82 previous reports of thermal instability of this enzyme 32 . The enzyme exhibited first-order 83 degradation with a half-life (t½) ...

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Section: Resultssupporting

confidence: 59%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Surpassing thermodynamic, kinetic, and stability barriers to isomerization catalysis for tagatose biosynthesis

Bober

Nair

2019

Preprint

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“…However, . are well documented for D-L. salivarius [7,9,14]. Henceforth, authors have considered to UJS 003 as a model strain to tagatose producing capability.…”

Section: Molecular Identification Of Bacterial Strainmentioning

confidence: 99%

“…Therefore, the production of D-tagatose by biological approaches is found to be more desired as safer regards to both health and environment. In recent years, many studies demonstrated the production of D-tagatose via the enzymatic catalysis by employing Larabinose isomerase which is capable of directly converting D-galactose to D-tagatose [6][7][8][9]. However, the key determinative steps are to screen the strains with high L-arabinose enzyme activity suitable for large-scale production, to construct the enzyme expression system and to explore the appropriate conversion process.…”

Section: Introductionmentioning

confidence: 99%

Screening and Identification of Lactic Acid Bacteria with D-tagatose Production Capability

Wen

Zhou

Huang

et al. 2019

BJI

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Aims: D-tagatose is a natural ketohexose which can be used as a functional sweetener in foods, diary and beverages products. Isolation of new bacterial strains having the ability to produce Dtagatose is a continuously trending topic of research. Study Design: Screening of strains with D-tagatose production by identification and determination of its ability to produce D-tagatose content. Place and Duration of Study: School of Food and Biological Engineering, Jiangsu University between May 2018 and April 2019. Methodology: We initially screened and identified the strains capable of producing D-tagatose through kimchi liquid, and determined the species and genetic characteristics of the strain by physiological, biochemical and molecular biological identification, and then determined the content of D-tagatose by high performance liquid chromatography. Finally calculate the ability of the strain to produce D-tagatose. Li et al.; BJI, 23(2): 1-6, 2019; Article no.BJI.49412 2 Results: In this study, 4 strains of lactic acid bacteria (LAB) were isolated from kimchi sample. The isolates were identified as Lactobacillus spp. (Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus salivarius) on the basis of morphological, physicochemical characteristics and analysis of 16S rDNA gene sequence. Because of the novelty, strain designated as L. salivarius UJS 003 was considered for D-tagatose yield. Fermentation of D-tagatose was carried out using galactose as substrate for 48 hr at 37 °C, and HPLC method was used to determine the yield. The experimental results exhibited a D-tagatose yield of 3.134 g/L by L. salivarius UJS 003. Conclusion: The strain UJS 003 represented as a potent D-tagatose producer and could be useful in a variety of biotechnological and industrial processes, particularly food and beverage industries. Short Research Article

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Production of _D‐tagatose, bioethanol, and microbial protein from the dairy industry by‐product whey powder using an integrated bioprocess

Ma,

Li,

et al. 2024

Biotechnology Journal

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We designed and constructed a green and sustainable bioprocess to efficiently coproduce D‐tagatose, bioethanol, and microbial protein from whey powder. First, a one‐pot biosynthesis process involving lactose hydrolysis and D‐galactose redox reactions for D‐tagatose production was established in vitro via a three‐enzyme cascade. Second, a nicotinamide adenine dinucleotide phosphate‐dependent galactitol dehydrogenase mutant, D36A/I37R, based on the nicotinamide adenine dinucleotide‐dependent polyol dehydrogenase from Paracoccus denitrificans was created through rational design and screening. Moreover, an NADPH recycling module was created in the oxidoreductive pathway, and the tagatose yield increased by 3.35‐fold compared with that achieved through the pathway without the cofactor cycle. The reaction process was accelerated using an enzyme assembly with a glycine–serine linker, and the tagatose production rate was 9.28‐fold higher than the initial yield. Finally, Saccharomyces cerevisiae was introduced into the reaction solution, and 266.5 g of D‐tagatose, 162.6 g of bioethanol, and 215.4 g of dry yeast (including 38% protein) were obtained from 1 kg of whey powder (including 810 g lactose). This study provides a promising sustainable process for functional food (D‐tagatose) production. Moreover, this process fully utilized whey powder, demonstrating good atom economy.

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d -Tagatose production by permeabilized and immobilized Lactobacillus plantarum using whey permeate

Cited by 49 publications

References 27 publications

Surpassing thermodynamic, kinetic, and stability barriers to isomerization catalysis for tagatose biosynthesis

Surpassing thermodynamic, kinetic, and stability barriers to isomerization catalysis for tagatose biosynthesis

Screening and Identification of Lactic Acid Bacteria with D-tagatose Production Capability

Production of _D‐tagatose, bioethanol, and microbial protein from the dairy industry by‐product whey powder using an integrated bioprocess

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d -Tagatose production by permeabilized and immobilized Lactobacillus plantarum using whey permeate

Cited by 49 publications

References 27 publications

Surpassing thermodynamic, kinetic, and stability barriers to isomerization catalysis for tagatose biosynthesis

Surpassing thermodynamic, kinetic, and stability barriers to isomerization catalysis for tagatose biosynthesis

Screening and Identification of Lactic Acid Bacteria with D-tagatose Production Capability

Production of D‐tagatose, bioethanol, and microbial protein from the dairy industry by‐product whey powder using an integrated bioprocess

Contact Info

Product

Resources

About

Production of _D‐tagatose, bioethanol, and microbial protein from the dairy industry by‐product whey powder using an integrated bioprocess