High Production of Trametes cinnabarina Laccase (lac1) by Suspended and Immobilized Cells of Recombinant Pichia pastoris from Crude Glycerol

Bhattacharyya, Aniket; Ahmed, Mostak; Wadhwa, Ridhima; Aggarwal, Sudhir; Mustafiz, Ananda; Rajagopalan, Gobinath

doi:10.1007/s12649-021-01661-1

Cited by 8 publications

(9 citation statements)

References 38 publications

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“…Furthermore, the enzymatic activity of Lac in the fermented broth was measured and found to be 18.4 ± 1.8 U mL −1 after 192 h of fermentation (day 8), surpassing the results reported by Cruz et al ., 3 who achieved a maximum activity of 4.9 U mL −1 for the same microorganism and substrate. Additionally, the observed Lac activity exceeded the results obtained by Bhattacharyya et al ., 5 who achieved a pure Lac production of 2.8 U mL −1 using a glycerol substrate.…”

Section: Resultscontrasting

confidence: 74%

“…This significant reduction in reducing sugar concentration indicates that the microorganism effectively utilized these carbohydrates for its growth and enzyme production, aligning with previous studies conducted by Fernandes et al 27 and Vilar et al 4 Furthermore, the enzymatic activity of Lac in the fermented broth was measured and found to be 18.4 ± 1.8 U mL −1 after 192 h of fermentation (day 8), surpassing the results reported by Cruz et al, 3 who achieved a maximum activity of 4.9 U mL −1 for the same microorganism and substrate. Additionally, the observed Lac activity exceeded the results obtained by Bhattacharyya et al, 5 who achieved a pure Lac production of 2.8 U mL −1 using a glycerol substrate. These findings highlight the potential of utilizing PW as a valuable resource for enzymatic production, and the results obtained demonstrate the improved efficiency of Lac production compared with previous studies.…”

Section: Production Of Laccontrasting

confidence: 74%

“…4 The promising oxidative potential of enzymes such as Lac, manganese peroxidase, lignin peroxidase, and versatile peroxidase in degrading lignocellulosic compounds as carbon sources offers advantages across various industrial sectors. 5 These enzymes have demonstrated their effectiveness in several industrial applications, including the textile industry, 6 food, pharmaceuticals, paper, and cellulose, 7 and for the degradation of emerging pollutants. 8 According to MarketWatch reports, 9 the global Lac market reached a value of US$2.96 billion at the end of 2025.…”

Section: Introductionmentioning

confidence: 99%

“…These enzymes can be obtained from the white‐rot basidiomycete fungus Pleurotus sajor‐caju , offering a potential solution for reducing the environmental impact associated with this effluent 4 . The promising oxidative potential of enzymes such as Lac, manganese peroxidase, lignin peroxidase, and versatile peroxidase in degrading lignocellulosic compounds as carbon sources offers advantages across various industrial sectors 5 …”

Section: Introductionmentioning

confidence: 99%

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Thermosensitive polymer‐assisted extraction and purification of fungal laccase from citrus pulp wash effluent

Santana,

Barros,

Canuto

et al. 2023

J Sci Food Agric

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BackgroundThis study explores the use of liquid‐liquid extraction (LLE) with thermosensitive polymers for producing laccase (Lac) from Pleurotus sajor‐caju. This process leverages liquid waste from the citrus industry, specifically pulp wash. The research delves into extractive fermentation and thermoseparation, both processes were facilitated by a polymer exhibiting a lower critical solution temperature (LCST) transition.ResultsKey factors considered include the choice of polymer, its concentration, pH, separation temperature, and the behavior of the polymer‐rich phase post‐extractive fermentation concerning LCST. Notably, under conditions of 45% by weight of Pluronic L‐61 and a pH of 5.0 at 25°C, the laccase was achieved an enhancement in the purification factor (PF) of 28.4‐fold, compared to the laccase obtained directly from the fermentation process on the 8th day. There was an 83.6% recovery of the laccase enzyme in the bottom phase of the system. Additionally, the unique properties of Pluronic L‐61, which can induce phase separation and also allow for thermoseparation, led to a secondary fraction (aqueous solution) of laccase with PF of 2.1 ± 0.1‐fold (at 32 ± 0.9 °C and 30 ± 0.3 min without stirring) from polymeric phase (top‐phase). The FTIR analysis validated the separation data, particularly highlighting the α‐helix content in the Amide I region (1600 ‐ 1700 cm‐1).ConclusionIn summary, the insights from this study pave the way for broader industrial applications of these techniques, underscoring benefits like streamlined process integration, heightened selectivity, and superior separation efficacy.This article is protected by copyright. All rights reserved.

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Section: Resultscontrasting

confidence: 74%

Section: Production Of Laccontrasting

confidence: 74%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Thermosensitive polymer‐assisted extraction and purification of fungal laccase from citrus pulp wash effluent

Santana,

Barros,

Canuto

et al. 2023

J Sci Food Agric

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“…A fungal laccase production process was designed by recycling the P. pastoris biomass and replacing pure glycerol with crude glycerol. 2,343 U.L −1 laccase was produced after 48 h. Recycling of free cells for 6 times produced > 8800 U laccase, while recycling of immobilized cells for 18 batches resulted with 27,681–33,926 U laccase production with pure/crude glycerol media [ 109 ].…”

Section: Introductionmentioning

confidence: 99%

Second generation Pichia pastoris strain and bioprocess designs

Ergün

Laçın

Çaloğlu

et al. 2022

Biotechnol Biofuels

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Yeast was the first microorganism used by mankind for biotransformation processes that laid the foundations of industrial biotechnology. In the last decade, Pichia pastoris has become the leading eukaryotic host organism for bioproduct generation. Most of the P. pastoris bioprocess operations has been relying on toxic methanol and glucose feed. In the actual bioeconomy era, for sustainable value-added bioproduct generation, non-conventional yeast P. pastoris bioprocess operations should be extended to low-cost and renewable substrates for large volume bio-based commodity productions. In this review, we evaluated the potential of P. pastoris for the establishment of circular bioeconomy due to its potential to generate industrially relevant bioproducts from renewable sources and waste streams in a cost-effective and environmentally friendly manner. Furthermore, we discussed challenges with the second generation P. pastoris platforms and propose novel insights for future perspectives. In this regard, potential of low cost substrate candidates, i.e., lignocellulosic biomass components, cereal by-products, sugar industry by-products molasses and sugarcane bagasse, high fructose syrup by-products, biodiesel industry by-product crude glycerol, kitchen waste and other agri-food industry by products were evaluated for P. pastoris cell growth promoting effects and recombinant protein production. Further metabolic pathway engineering of P. pastoris to construct renewable and low cost substrate utilization pathways was discussed. Although, second generation P. pastoris bioprocess operations for valorisation of wastes and by-products still in its infancy, rapidly emerging synthetic biology tools and metabolic engineering of P. pastoris will pave the way for more sustainable environment and bioeconomy. From environmental point of view, second generation bioprocess development is also important for waste recycling otherwise disposal of carbon-rich effluents creates environmental concerns. P. pastoris high tolerance to toxic contaminants found in lignocellulosic biomass hydrolysate and industrial waste effluent crude glycerol provides the yeast with advantages to extend its applications toward second generation P. pastoris strain design and bioprocess engineering, in the years to come. Graphical Abstract

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High level of 1,3‐propanediol production from crude glycerol by Clostridium strain BOH3: critical roles of inoculum and substrate concentration

Rajagopalan,

Jain,

Poosarla

et al. 2024

J of Chemical Tech & Biotech

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BACKGROUND1,3‐Propanediol (PDO) is a raw material/specialty chemical/additive in several industrial applications. A new wild‐type Clostridium strain BOH3 can utilize crude glycerol (CG) and produce PDO. This investigation reveals its potential for PDO production from CG.RESULTSInitially, inoculum of strain BOH3 was cultured from reinforced clostridial medium (RCM) with glucose (medium‐A), and modified‐RCM with glycerol (medium‐B). While fermenting 20 g/L glycerol, the cells cultivated from medium‐A produced 5.75 g/L PDO (as major product) with 3.10 g/L butanol (byproduct), whereas the cells cultivated from medium‐B produced 11.01 g/L PDO with 2.50 g/L butyric, and 1.01 g/L acetic acids, and no butanol was produced. To enhance PDO production from the batch‐process, various parameters including pure glycerol/CG concentration (80 g/L), inoculum age (15 h) and size (12.50%v/v) of cells from medium‐B, initial medium‐pH (pH 6.4), and incubation temperature (39 °C) were studied; the optimal values obtained are given in their respective parenthesis. Under optimal conditions, PDO production was 42.58–47.15 g/L (0.65–0.79 mol‐PDO/mol‐glycerol) from CG‐based rich/mineral media. Furthermore, fed‐batch fermentation with 50–80 g/L initial CG‐concentration was tested to enhance the PDO production. Among these, 60 g/L initial CG‐concentration assisted strain BOH3 in utilizing a high level of CG (~174 g/L); it supported a higher‐titer of PDO production (103.55–116.45 g/L), with the highest yield (0.72–0.81 mol‐PDO/mol‐glycerol) ever reported.CONCLUSIONClostridium strain BOH3 shows the potential to support a high titer (>116 g/L) with the highest yield (>0.80 mol/mol) of PDO production from CG (a waste). It can be considered for industrial‐scale PDO production.This article is protected by copyright. All rights reserved.

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High Production of Trametes cinnabarina Laccase (lac1) by Suspended and Immobilized Cells of Recombinant Pichia pastoris from Crude Glycerol

Cited by 8 publications

References 38 publications

Thermosensitive polymer‐assisted extraction and purification of fungal laccase from citrus pulp wash effluent

Thermosensitive polymer‐assisted extraction and purification of fungal laccase from citrus pulp wash effluent

Second generation Pichia pastoris strain and bioprocess designs

High level of 1,3‐propanediol production from crude glycerol by Clostridium strain BOH3: critical roles of inoculum and substrate concentration

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