Decolorization of Direct Blue GLL with enhanced lignin peroxidase enzyme production in Comamonas sp UVS

BACKGROUND: The present work aims to study the production of lignin peroxidase (LiP) enzyme by Comamonas sp UVS using various media, and lignocellulosic waste materials, and its effect on decolorization of Direct Blue GLL (DBGLL). RESULTS: Yeast extract medium was found to be more effective for the production of LiP and also for the decolorization of DBGLL. The bagasse powder along with yeast extract induced LiP activity. Comamonas sp UVS decolorized DBGLL dye (50 mg L^?1) within 13 h at static condition in YE broth. It could degrade up to 300 mg L^?1 of dye within 55 h. The maximum rate (Vmax) of decolorization was 12.41 ± 0.55 mg dye g cell^?1 h^?1 with the Michaelis constant (Km) value as 6.20 ± 0.27 mg L^?1. The biodegradation was monitored by UV‐Vis, GC‐MS and HPLC. CONCLUSION: The use of agricultural by‐products for the activity enhancement of the ligninolytic enzymes is a cost effective process. It also resolves the problem of the disposal of agro‐residues. This system can be applied for the degradation of different recalcitrant compounds. Copyright © 2008 Society of Chemical Industry     

The use of olive mill wastewater by wild type Yarrowia lipolytica strains: medium supplementation and surfactant presence effect

BACKGROUND: The aim of this work was to study the ability of two different wild type strains of the yeast Yarrowia lipolytica to grow on olive mill wastewater (OMW) and their potential to produce high‐value products such as lipases. Factors that affect cellular growth and OMW degradation were studied, such as nitrogen supplementation, cells concentration and surfactant addition. RESULTS: Both strains, W29 and IMUFRJ 50682, were able to grow in OMW with 19 g L^?1 of COD and approximately 800 mg L^?1 of total phenols concentration. The strain W29 presented the highest potential for extracellular lipase production in OMW medium. Lipase productivity was improved by the medium supplementation with ammonium sulphate up to 6 g L^?1, leading to 80% of COD degradation and 70% of total phenols reduction. The surfactant Tween 80 enhanced cell growth and COD degradation, but had a negative effect on lipase activity. CONCLUSIONS Y. lipolytica has a great potential for OMW valorisation by its use as culture medium for biomass and enzymes production. The operating conditions that favoured lipase production differ from the conditions that improve COD reduction. Copyright © 2008 Society of Chemical Industry     

Removal of lignin in a liquid system by an isolated fungus

The screening of a strain which could perform lignin removal was carried out. Based on taxonomic study the isolated strain (LM‐2) was identified as Penicillium sp. LM‐2 could decolorize 0.6 g dm^?3 lignin within 4 days in a shaking culture at 25 °C. The efficiency of decolorization of the lignin was over 80% in the pH range of 4.0–6.0, but was low above pH 6.2. The rise of temperature had a slight adverse effect on the lignin decolorization in the range of 25–35 °C. Lignolytic enzymes such as lignin peroxidase, manganese peroxidase and laccase were not detected in the culture broth or within the fungal cells. The lignin was removed from the high molecular weight fraction mainly by adsorption and accumulation inside the cells. © 2001 Society of Chemical Industry     

L(+)‐Lactic acid production using poly(vinyl alcohol)‐cryogel‐entrapped Rhizopus oryzae fungal cells

A new immobilized biocatalyst based on Rhizopus oryzae fungal cells entrapped in poly(vinyl alcohol)‐cryogel was evaluated in both the batch and semi‐batch processes of L (+)‐lactic acid (LA) production, when glucose, acid hydrolysates of starch or gelatinized potato starch were used as the main substrates. Under the batch conditions, the immobilized biocatalyst developed produced LA with yields of 94% and 78% from glucose and acid starch hydrolysates, respectively. Semi‐batch conditions enabled product yields of 52% and 45% to be obtained with the corresponding substrates. The highest process productivity (up to 173 g L^?1) was reached under semi‐batch conditions. Potato starch (5–70 g L^?1) was also transformed into lactic acid by immobilized R. oryzae. It was shown that long‐term operation of the immobilized biocatalyst (for at least 480 h) produced a low decrease in metabolic activity. Copyright © 2006 Society of Chemical Industry     

Optimization of extraction of bulk enzymes from spent mushroom compost

The profiling of ligninase, hemicellulase and cellulase of Pleurotus sajor‐caju after inoculation of spawn in bags containing sawdust was done at monthly intervals for a period of 6 months. Xylanase (EC 3.2.1.8) was produced throughout the 6 months studied with the productivity range from 5.60 to 7.51 U g^?1. Cellulase (EC 3.2.1.4) and β‐glucosidase (EC 3.2.1.21) productivities were highest at 4 months, producing 3.31 U g^?1 and 121.13 U g^?1 respectively. Laccase (EC 1.10.3.2) productivity was highest at 2 months with a value of 7.59 U g^?1. Lignin peroxidase (EC 1.11.1.14) productivity was highest at 5 months with a value of 206.20 U g^?1. Total soluble proteins were highest at 4 months with a value of 0.139 mg cm^?3. The profiling of lignin peroxidase in 5‐month‐old spent mushroom compost was monitored over a period of 10 months. It was observed that lignin peroxidase was produced throughout the period but productivity was variable. The average lignin peroxidase productivity ranged from 30 to 110 U g^?1. The activities of the enzymes extracted in tap water at pH 8.4 were comparable to that extracted in 50 mmol sodium citrate buffer at pH 4.8 and distilled water at pH 5.2 at 4 °C using an incubator shaker at 200 rpm for 18 h. The optimum extraction time was 1 h using an incubator shaker at 4 °C. When an incubator shaker was used, there was no significant difference in the recovery of xylanase, cellulase and laccase at different pH values at 4 °C and 28 °C. No significant difference was observed in the recovery of β‐glucosidase using an incubator shaker at different pH values at 4 °C although the enzyme recovery was slightly higher at pH 8.12, with a value of 29.27 U g^?1. The optimum extraction of β‐glucosidase was at pH 4 at room temperature using an incubator shaker. For the lignin peroxidase enzyme, the optimum pH for extraction was 6 at 4 °C and pH 7 at room temperature using an incubator shaker at 200 rpm for 1 h. Homogenization for 8 min at 8000 rpm using tap water at pH 4 had an advantage over the use of the incubator shaker for the extraction as high titers of enzymes were recovered. Copyright © 2003 Society of Chemical Industry     

Purification and enzymatic properties of the extracellular and constitutive chitosanase produced by Bacillus subtilis RKY3

BACKGROUND: Purification and enzymatic properties of a chitosanase from Bacillus subtilis RKY3 have been investigated to produce a chitooligosaccharide. The enzyme reported was extracellular and constitutive, which was purified by two sequential steps including ammonium sulfate precipitation and ion exchange chromatography. RESULTS: Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of the purified chitosanase revealed one single band corresponding to a molecular weight of around 24 kDa. The highest chitosanase activity was found to be at pH 6.0 and at 60 °C. Although the mercaptide forming agents such as Hg²⁺ (10 mmol L^?1) and p‐hydroxymercuribenzoic acid (1 mmol L^?1, 10 mmol L^?1) significantly or totally inhibited the enzyme activity, its activity was enhanced by the presence of 10 mmol L^?1 Mn²⁺. The enzyme showed activity for hydrolysis of soluble chitosan and glycol chitosan, but colloidal chitin, carboxymethyl cellulose, crystalline cellulose, and soluble starch were not hydrolyzed. The analysis of chitosan hydrolysis by thin‐layer chromatography and viscosity variation revealed that the purified enzyme should be endosplitting‐type chitosanase. CONCLUSION: The chitosanase produced by Bacillus subtilis RKY3 was a novel chitosanlytic enzyme with relatively low molecular weight, which is a versatile enzyme for chitosan hydrolysis because it could hydrolyze soluble chitosan into a biofunctional oligosaccharide at a high level. Copyright © 2011 Society of Chemical Industry     

Optimization of alkaline protease production from Shewanella oneidensis MR‐1 by response surface methodology

BACKGROUND: The proteases are among the most important groups of enzymes. Therefore, it is important to produce inexpensive and optimized media for large‐scale commercial production. In the present work, three different Shewanella species were screened on skim milk agar medium for their ability to produce alkaline protease. The effects of different culture conditions were optimized for alkaline protease production by S. oneidensis MR‐1 using a Box–Behnken design combined with response surface methodology (RSM). RESULTS: Highest yield (112.90 U mL^?1) of protease production was obtained at pH 9.0, a temperature of 30 °C, glucose (12.5 g L^?1), tryptone (12.5 g L^?1) and an incubation period of 36 h. A second‐order polynomial regression model was used for analysis of the experiment. The experimental values were in good agreement with predicted values, with correlation coefficient 0.9996. CONCLUSION: Carbon and nitrogen, pH, temperature and incubation period were chosen as the main factors to be used in an experimental design for optimization to produce low‐cost enzymes, potentially for use on an industrial scale. A 60% increase in enzyme activity was achieved in the optimized medium compared with the original medium. Copyright © 2008 Society of Chemical Industry     

Enhanced saccharification of corn straw pretreated by alkali combining crude ligninolytic enzymes

BACKGROUND: This work investigated the monokaryogenesis of dikaryon strains of Trametes hirsuta by protoplasts regeneration for extracellular ligninolytic enzyme production. Saccharification of corn straw was enhanced by alkali pretreatment combining crude ligninolytic enzymes. RESULTS: Effectiveness of alkali pretreatment of corn straw on delignification was evaluated under different concentrations. About 45% lignin loss was achieved at the concentration of 1.5% NaOH. In addition, 79.0% sugar yield was obtained after combined pretreatment with NaOH and crude ligninolytic enzyme produced from monokaryotic strains of Trametes hirsuta. Scanning electron microscopy (SEM) images showed that the porosity and surface area increased significantly after combined pretreatment. The FTIR spectra indicated that great intensity changes occurred at the 890–900 cm^?1, 1509–1513 cm^?1 and 1595 cm^?1 bands. CONCLUSION: The proposed combined pretreatment removes lignin and enhances saccharification of corn straw effectively. Copyright © 2012 Society of Chemical Industry     

Statistical Optimization of Culture Conditions and Characterization for Ligninolytic Enzymes Produced from Rhizopus Sp. Using Prickly Palm Cactus Husk

Dry prickly palm cactus (Nopalea cochenillifera) husk was investigated as a substrate for Rhizopus sp. cultivation in the solid state, aiming at the production of laccase (Lac), lignin peroxidase (LiP), and manganese peroxidase (MnP). The optimization of fermentation was evaluated by an experimental design and it was obtained, for each enzyme, maximum productivities (U g^?1 h^?1) of: 0.085 ± 0.02 (MnP), 0.066 ± 0.001 (LiP), and 0.023 ± 2.3.10^?4 (Lac), at the conditions of 10 g of substrate, 72 h of fermentation, a_w = 0.865, and 30°C. The enzymes thermal and pH stabilities were evaluated and it was observed better results at temperatures no higher than 60°C and pH of 5.0; in addition, the storage of these enzymes was better at ? 25°C than at 4°C. Since the prickly palm cactus is an agricultural substrate and specially because of its low cost, it is important to propose different applications for it as, for example, an alternative substrate for biotechnological processes.     

Black liquor detoxification by laccase of Trametes versicolor pellets

Black liquors from a soda pulping mill were treated with the white‐rot fungus Trametes versicolor to detoxify and reduce colour, aromatic compounds and chemical oxygen demand (COD). The fungus was used in the form of pellets in aerated reactors (fluidized, stirred and air‐pulsed reactors). Reductions in colour and aromatic compounds of 70–80% and in COD of 60% were achieved. During the different experiments, laccase activity was detected but neither lignin peroxidase (LiP) nor manganese peroxidase activities were detected, although T versicolor is able to produce these enzymes. Experiments also showed a LiP activity inhibitory effect produced by lignin. From the results obtained, it can be concluded that there is a relationship between laccase production and toxicity reduction. This correlation responds to the equation Laccase production = 1.57 LN (toxicity reduction) ?16.40. Copyright © 2003 Society of Chemical Industry     

Removal of aqueous phenolic compounds from a model system by oxidative polymerization with turnip (Brassica napus L var purple top white globe) peroxidase

Turnip roots, which are readily available in Mexico, are a good source of peroxidase, and because of their kinetic and biochemical properties have a high potential as an economic alternative to horseradish peroxidase (HRP). The efficiency of using turnip peroxidase (TP) to remove several different phenolic compounds as water‐insoluble polymers from synthetic wastewater was investigated. The phenol derivatives studied included phenol, 2‐chlorophenol, 3‐chlorophenol, o‐cresol, m‐cresol, 2,4‐dichlorophenol and bisphenol‐A. The effect of pH, substrate concentration, amount of enzyme activity, reaction time and added polyethylene glycol (PEG) was investigated in order to optimize reaction conditions. A removal efficiency ≥85% was achieved for 0.5 mmol dm^?3 phenol derivatives at pH values between 4 and 8, after a contact time of 3 h at 25 °C with 1.28 U dm^?3 of TP and 0.8 mmol dm^?3 H₂O₂. Addition of PEG (100–200 mg dm^?3) significantly reduced the reaction time required (to 10 min) to obtain >95% removal efficiency and up to 230% increase in remaining TP activity. A relatively low enzyme activity (0.228 U dm^?3) was required to remove >95% of three phenolic solutions in the presence of 100–200 mg dm^?3 PEG. TP showed efficient and fast removal of aromatic compounds from synthetic wastewaters in the presence of hydrogen peroxide and PEG. These results demonstrate that TP has good potential for the treatment of phenolic‐contaminated solutions. © 2002 Society of Chemical Industry     

Inoculation of Filamentous Fungi in Manufactured Gas Plant Site Soils and PAH Transformation

Solid carriers have been developed to inoculate Trametes versicolor and Cunninghamella elegans into manufactured gas plant site soils. Pelleted wheat bran carriers were very efficient in stimulating the growth of fungi in an industrial soil containing about 2800 mg kg^?1 PAHs. Fungal biomass and activity of extracellular laccases, enzymes produced by T. versicolor as markers of metabolic activity in the contaminated soil, both decreased after 2 weeks of incubation. Supplementing the soil with a mixture of carbon, nitrogen and phosphorus enhanced the fungal activity period. A 38% decrease of solvent extractable PAHs was observed in manufactured gas plant site soils when supplemented with T. versicolor, Glucidex 19^TM, ammonium nitrate, lime phosphate and Montanox 80^TM, after 20 weeks. Then, the degradation proceeded more slowly during the following 30 weeks, and reached 43% of initial extractable PAHs. Some factors governing a limited PAH biotransformation in the soil are discussed.     

Preparation of a promising whole cell biocatalyst of Geotrichum sp. lipase and its properties

BACKGROUND: As a new protein expression and self‐immobilization system, cell‐surface displayed enzymes have attracted increasing attention. In this study, Geotrichum sp. lipase (GSL), an important enzyme for the enrichment of polyunsaturated fatty acids (PUFAs), was first displayed on the cell surface of Saccharomyces cerevisiae. RESULTS: The activity of displayed GSL was higher (43.7 U g^?1 dry cell) than that of Candida antarctica lipase B (26.26 U g^?1 dry cell) and that of Rhizopus oryzae lipase (4.1 U g^?1 dry cell). It also exhibited higher thermostability than the free lipase, and retained 89% of the original activity after incubation at 40 °C for 3 h, compared with 48% at 35 °C for the free lipase at pH 8.5. Interestingly, the displayed lipase had a wider pH range and better pH stability. It had higher activity at all pH values than the free GSL, and retained 86% of the original activity in the pH range 9.5 to 10.5, whereas the activity of the free GSL could not be detected at pH 10. CONCLUSION: This work presented a method to prepare a whole‐cell biocatalyst with better stability and broader pH tolerance which will provide a useful strategy for other cost‐effective self‐immobilized industrial lipases. Copyright © 2011 Society of Chemical Industry     

D‐arabitol production from lactose by Kluyveromyces lactis at different aerobic conditions

BACKGROUND: The pentitol D‐arabitol has been produced from D‐glucose utilizing osmophilic yeast strains, however, there are remarkably few reports available on the production of D‐arabitol from lactose. Previous studies in the laboratory have shown that the osmophilic yeast Kluyveromyces lactis NBRC 1903 can convert lactose to extracellular D‐arabitol without extracellular accumulation of D‐glucose or D‐galactose. The present study was undertaken to determine the participation of aeration on the D‐arabitol synthesis in K. lactis NBRC 1903. RESULTS: The highest D‐arabitol concentration of 91.7 mmol L^?1 was achieved after 120 h cultivation in medium containing 555 mmol L^?1 of lactose with initial volumetric liquid‐phase mass transfer coefficient of oxygen (k_La)₀ of 85.5 h^?1. The fractional yield of D‐arabitol was affected by not only aeration but also growth phase. The highest fractional yield of D‐arabitol in terms of lactose consumption was 0.255 that was obtained at stationary phase with (k_La)₀ of 85.5 h^?1. CONCLUSION: It was found that oxygen supply is a key factor in the production of D‐arabitol. Patterns of metabolism were classified according to the level of oxygen supply and the growth phase. Copyright © 2010 Society of Chemical Industry     

Kinetic data and substrate specificity of a keratinase from Chryseobacterium sp. strain kr6

BACKGROUND: Keratinases are important enzymes for biotechnological processes involving keratin hydrolysis. In this work substrate specificity and kinetic properties of a keratinase from Chryseobaterium sp. were investigated. RESULTS: The optimal conditions for activity of purified keratinase with respect to pH, temperature and sodium chloride concentration were established using factorial design and surface response techniques. The optimum conditions for keratinase activity were pH from 7.4 to 9.2, temperature from 35 °C to 50 °C and NaCl concentration from 50 to 340 mmol L^?1, having azocasein as substrate. Subsequently, the kinetic parameters for this substrate were determined to be K_m = 0.75 mg mL^?1 and V_max = 59.5 U min^?1. The K_i value for 1,10‐phenanthroline was estimated at 0.78 mmol L^?1. The enzyme specificity was evaluated over different synthetic and insoluble substrates. The protease exhibited specificity with selectivity for hydrophobic and positively charged residues. In relation to the insoluble substrates, the enzyme hydrolyzed preferably chicken nails. CONCLUSIONS: This enzyme effectively hydrolyzes insoluble keratin substrates. The knowledge of keratinase properties is an essential step in the development of biotechnological processes involving keratin hydrolysis. Copyright © 2008 Society of Chemical Industry     

Production of organic solvent tolerant lipase by Staphylococcus caseolyticus EX17 using raw glycerol as substrate

BACKGROUND: In this work we used Plackett–Burman statistical design and central composite design in order to optimize culture conditions for lipase production by Staphylococcus caseolyticus strain EX17 growing on raw glycerol, which was obtained as a by‐product of the enzymatic synthesis of biodiesel. The stability of lipase was verified over several organic solvents, such as methanol, ethanol and n‐hexane. RESULTS: Optimal culture conditions for lipase production were found to be 36 °C, initial pH 8.12, glycerol 30 g L^?1, olive oil 3.0 g L^?1, and soybean oil 2.5 g L^?1, with 145.8 U L^?1 of enzyme activity. When commercial glycerol was substituted by the raw glycerol from biodiesel synthesis, lipolytic activity was 127.3 U L^?1. Experimental validation of enzyme production matched values predicted by the mathematical model, which was 138.3 U L^?1. Stability tests showed that lipase from S. caseolyticus EX17 was stable in methanol, ethanol, and n‐hexane. CONCLUSIONS: Results obtained in this work suggest that raw glycerol can be used for lipase production by S. caseolyticus EX17 and that this enzyme has a potential application in the synthesis of biodiesel. Copyright © 2008 Society of Chemical Industry     

Utilization of lignocellulosic waste for ethanol production : Enzymatic digestibility and fermentation of pretreated shea tree sawdust

Enzymatic hydrolysis and fermentation methods were evaluated on alkaline peroxide pretreated shea tree sawdust conversion to ethanol. Optimum pretreatment conditions of 120 °C reaction temperature, 30 min reaction time, and 20 mL L^?1 of water hydrogen peroxide concentration (1%(v/v)H₂O₂) solubilized 679 g kg^?1 of hemicellulose and 172 g kg^?1 of lignin. 617 g kg^?1 cellulose was retained in the solid fraction. The maximum yield of reducing sugar with optimized enzyme loadings by two enzyme preparations (cellulase and β-glucosidase) was 165 g kg^?1 of dry biomass. The ethanol yield was 7.35 g L^?1 after 72 h incubation period under the following conditions: 2% cellulose loading, enzyme concentration was 25 FPU (g cellulose)^?1 loading, yeast inoculums was 10% (v/v), 32 oC, and pH 4.8. The pretreatments gave information about the hindrances caused by lignin presence in lignocellulosic materials and that hemicelluloses are better hydrolyzed than lignin, thereby enhancing enzymatic digestibility of the sawdust material.     

Fermentation and oxygen transfer characteristics in recombinant human growth hormone production by Pichia pastoris in sorbitol batch and methanol fed‐batch operation

BACKGROUND: The influence of methanol feed rate on recombinant human growth hormone (rhGH) production by Pichia pastoris hGH‐Mut⁺ in medium containing sorbitol was investigated at three different specific growth rates (µ), namely, 0.02 (MS‐0.02), 0.03 (MS‐0.03), and 0.04 (MS‐0.04). RESULTS: Increasing methanol feed rate above MS‐0.03 did not affect sorbitol consumption, showing that µ = 0.03 h^?1 is a threshold limiting value, above which sorbitol utilization became independent of methanol feed rate. Moreover, when sorbitol was consumed, no further cell growth was observed. Increase in methanol feed rate triggered cell synthesis and the highest cell concentration was obtained at MS‐0.04 as 48 g L^?1 (t = 18 h); whereas, the highest rhGH production, 270 mg L^?1, was obtained at MS‐0.03 as a consequence of lower extracellular protease production and higher AOX activity (41 U g^?1 CDW). Oxygen uptake rate increased with increasing µ, having the maximum value, 76.6 mmol m^?3 s^?1, at MS‐0.04. K_La had a tendency to increase with µ, having a maximum value of 0.15 s^?1 at MS‐0.04 (t = 15 h). CONCLUSION: By considering rhGH concentration and oxygen transfer characteristics, the bioprocess can be improved by a two‐stage feeding strategy, operating at MS‐0.04 at the beginning of fermentation, and thereafter shifting to MS‐0.03. Copyright © 2009 Society of Chemical Industry     

Production of L‐methionine by immobilized pellets of Aspergillus oryzae in a packed bed reactor

Production of L ‐methionine by immobilized pellets of Aspergillus oryzae in a packed bed reactor was investigated. Based on the determination of relative enzymatic activity in the immobilized pellets, the optimum pH and temperature for the resolution reaction were 8.0 and 60 °C, respectively. The effects of substrate concentration on the resolution reaction were also investigated and the kinetic constants (K_m and V_m) of immobilized pellets were found to be 7.99 mmol dm^?3 and 1.38 mmol dm^?3 h^?1, respectively. The maximum substrate concentration for the resolution reaction without inhibition was 0.2 mol dm^?3. The L ‐methionine conversion rate reached 94% and 78% when substrate concentrations were 0.2 and 0.4 mol dm^?3, respectively, at a flow rate of 7.5 cm³ h^?1 using the small‐scale packed bed reactor developed. The half‐life of the L ‐aminoacylase in immobilized pellets was 70 days in continuous operation. All the results obtained in this paper exhibit a practical potential of using immobilized pellets of Aspergillus oryzae in the production of L ‐methionine. © 2002 Society of Chemical Industry     

Physicochemical characterization of lignin fractions sequentially isolated from bamboo (Dendrocalamus brandisii) with hot water and alkaline ethanol solution

Nine lignin fractions from bamboo (Dendrocalamus brandisii) were sequentially isolated with hot water at 80, 100, and 120°C for 3 h and 60% aqueous ethanol containing 0.25, 0.5, 1.0, 2.0, 3.0, and 5.0% NaOH at 80°C for 3 h. Molecular weight and purity analysis revealed that the lignin fractions isolated by hot water (L₁, L₂, and L₃) had lower weight-average molecular weights (between 1350 and 1490 g mol⁻¹) and contained much higher amounts of associated hemicelluloses (between 9.26 and 22.29%), while the lignin fractions isolated by alkaline aqueous ethanol (L₄, L₅, L₆, L₇, L₈, and L₉) had higher weight-average molecular weights (between 2830 and 3170 g mol⁻¹) and contained lower amounts of associated hemicelluloses (between 0.63 and 1.66%). Spectroscopy (UV, FTIR, ¹³C-NMR, and HSQC) analysis showed that the bamboo (Dendrocalamus brandisii) lignin was typical grass lignin, consisting of p-hydroxyphenyl (H), guaiacyl (G), and syringyl (S) units. The major interunit linkages presented in the alkaline aqueous ethanol extractable bamboo lignin were β-O-4′ aryl ether linkages (about 74.3%), followed by β-β′ resinol-type linkages and β-1′ spirodienone-type linkages (both for 7.8%), together with small amounts of β-5′ phenylcoumaran (6.8%) and p-hydroxycinnamyl alcohols end groups (3.1%). In addition, a small percentage (1.0%) of the lignin side-chain was found to be acetylated at the γ-carbon, predominantly over syringyl units. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012