Thermophilic Bacillus Coagulans Research Articles

Fermentative valorisation of xylose-rich hemicellulosic hydrolysates from agricultural waste residues for lactic acid production under non-sterile conditions

Lactic acid (LA) is a platform chemical with diverse industrial applications. Presently, commercial production of LA is dominated by microbial fermentation using sugary or starch-based feedstocks. Research pursuits emphasizing towards sustainable production of LA using non-edible and renewable feedstocks have accelerated the use of lignocellulosic biomass (LCB). The present study focuses on the valorisation of xylose derived from sugarcane bagasse (SCB) and olive pits (OP) through hydrothermal and dilute acid pretreatment, respectively. The xylose-rich hydrolysate obtained was used for LA production by homo-fermentative and thermophilic Bacillus coagulans DSM2314 strain under non-sterile conditions. The fed-batch mode of fermentation resulted in maximum LA titers of 97.8, 52.4 and 61.3 g/L with a yield of 0.77, 0.66 and 0.71 g/g using pure xylose, xylose-rich SCB and OP hydrolysates, respectively. Further, a two-step aqueous two-phase system (ATPS) extraction technique was employed for the separation and recovery of LA accumulated on pure and crude xylose. The LA recovery was 45 – 65% in the first step and enhanced to 80–90% in the second step.The study demonstrated an efficient integrated biorefinery approach to valorising the xylose-rich stream for cost-effective LA production and recovery.

Pilot Scale for Production and Purification of Lactic Acid from Ceratonia siliqua L. (Carob) Bagasse

The bioconversion of lignocellulose and organic waste bagasse to lactic acid (LA) is an important alternative process requiring valorization as a potentially viable method in the production of pure LA, to be utilized for various purposes. Carob (Ceratonia siliqua L.) biomass was used for the production of LA, using a thermophilic Bacillus coagulans isolate, cultivated in a batch pilot scale of 35 L fermenters without yeast extract supplementation, and operated for 50 h. During the fermentation process, most of the degradable sugar was consumed within 35 h and resulted in the production of 46.9 g/L LA, with a calculated LA yield of 0.72 g/g sugars and productivity at the log phase of 1.69 g/L/h. The use of LA for different industrial applications requires high purity; therefore, a downstream process (DSP) consisting of different purification stages was used, enabling us to reach up to 99.9% (w/w) product purity, which indicates that the process was very effective. The overall almost pure L-LA yield of the DSP was 56%, which indicates that a considerable amount of LA (46%) was lost during the different DSP stages. This is the first study in which carob biomass bagasse has been tested on a pilot scale for LA production, showing the industrial feasibility of the fermentation process.

High-Level fermentative production of Lactic acid from bread waste under Non-sterile conditions with a circular biorefining approach and zero waste discharge

Bread waste (BW) is a severe solid waste management problem in Europe. The current study demonstrates an environment-friendly solution by valorising BW into lactic acid (LA) and the corresponding solid residues generated during hydrolysis and fermentation to biogas. To this end, BW was saccharified through acidic and enzymatic hydrolysis, and the hydrolysate obtained was used for LA fermentation under non-sterile conditions using thermophilic Bacillus coagulans DSM1. Maximum glucose concentration achieved during acid hydrolysis with 2% (v/v) acid loading and 20% (w/v) solid loading was 67.9 g/L glucose, with a yield of 0.34 g/g BW. The LA accumulated with concentrated BW acid hydrolysate was 102.4 g/L with yield and productivity of 0.75 g/g and 1.42 g/L. h, respectively. For enzymatic hydrolysis, three commercial amylase preparations (Amyloglucosidase, Spirizyme, Dextrozyme) were employed. The highest glucose release (98.6 g/L) and yield (0.49 g glucose/g BW) was attained with Dextrozyme from Novozymes. The fed-batch fermentation by B. coagulans was conducted, using commercial glucose and glucose-rich BW hydrolysate from Dextrozyme. The LA titer, yield and productivity obtained with pure glucose were 222.7 g/L, 0.92 g/g and 1.86 g/L.h, respectively, whereas BW hydrolysate (BWH) resulted in 155.4 g/L LA, with a conversion yield and productivity of 0.85 g/g glucose and 1.30 g/L. h, respectively. Further to the LA biosynthesis, the solid residues generated during hydrolysis and fermentation were subjected to biogas generation, resulting in 553 mL CH4/g volatile solids under batch mode. This massive LA titer amassed under non-sterile conditions and integrated biogas production using fermented residues demonstrates a high potential for an integrated biorefinery based on BW.

Efficient Co-Utilization of Biomass-Derived Mixed Sugars for Lactic Acid Production by Bacillus coagulans Azu-10

Lignocellulosic and algal biomass are promising substrates for lactic acid (LA) production. However, lack of xylose utilization and/or sequential utilization of mixed-sugars (carbon catabolite repression, CCR) from biomass hydrolysates by most microorganisms limits achievable titers, yields, and productivities for economical industry-scale production. This study aimed to design lignocellulose-derived substrates for efficient LA production by a thermophilic, xylose-utilizing, and inhibitor-resistant Bacillus coagulans Azu-10. This strain produced 102.2 g/L of LA from 104 g/L xylose at a yield of 1.0 g/g and productivity of 3.18 g/L/h. The CCR effect and LA production were investigated using different mixtures of glucose (G), cellobiose (C), and/or xylose (X). Strain Azu-10 has efficiently co-utilized GX and CX mixture without CCR; however, total substrate concentration (>75 g/L) was the only limiting factor. The strain completely consumed GX and CX mixture and homoferemnatively produced LA up to 76.9 g/L. On the other hand, fermentation with GC mixture exhibited obvious CCR where both glucose concentration (>25 g/L) and total sugar concentration (>50 g/L) were the limiting factors. A maximum LA production of 50.3 g/L was produced from GC mixture with a yield of 0.93 g/g and productivity of 2.09 g/L/h. Batch fermentation of GCX mixture achieved a maximum LA concentration of 62.7 g/L at LA yield of 0.962 g/g and productivity of 1.3 g/L/h. Fermentation of GX and CX mixture was the best biomass for LA production. Fed-batch fermentation with GX mixture achieved LA production of 83.6 g/L at a yield of 0.895 g/g and productivity of 1.39 g/L/h.

Expeditious production of concentrated glucose-rich hydrolysate from sugarcane bagasse and its fermentation to lactic acid with high productivity

Sugarcane bagasse (SCB) is anticipated to emerge as a potential threat to waste management in India on account of cheap surplus energy options and lower incentives through its co-generation. Through biotechnological intervention, the efficient utilization of SCB is seen as an opportunity. The present study aimed towards expeditious production of concentrated glucose-rich hydrolysate from SCB. Alkali pretreated biomass was chosen for hydrolysis with a new generation cellulase cocktail, Cellic CTec2 dosed at 25 mg g−1 glucan content. A two-step (9% + 9%) substrate feeding strategy was adopted with a gap of an hour, and saccharification was terminated in three different ways. Irrespective of the methods employed for termination, ∼84.5% cellulose was hydrolyzed releasing ≥100 g L−1 glucose from 18% biomass. Direct use of glucose-rich filtrates yielded 69.2 ± 2.5 g L−1 of L (+) lactic acid using thermophilic Bacillus coagulans NCIM 5648. The best-attained glucose and lactic acid productivities during separate hydrolysis and fermentation (SHF) in the present study were 5.27 and 2.88 g L−1 h−1, respectively. A green and sustainable process is demonstrated for the production of industrially relevant sugars from SCB at high productivity and its valorization to bio-based lactic acid.

L-(+)-Lactic Acid from Reed: Comparing Various Resources for the Nutrient Provision of B. coagulans

Biotechnological production of lactic acid (LA) is based on the so-called first generation feedstocks, meaning sugars derived from food and feed crops such as corn, sugarcane and cassava. The aim of this study was to exploit the potential of a second generation resource: Common reed (Phragmites australis) is a powerfully reproducing sweet grass which grows in wetlands and creates vast monocultural populations. This lignocellulose biomass bears the possibility to be refined to value-added products, without competing with agro industrial land. Besides utilizing reed as a renewable and inexpensive substrate, low-cost nutritional supplementation was analyzed for the fermentation of thermophilic Bacillus coagulans. Various nutritional sources such as baker’s and brewer’s yeast, lucerne green juice and tryptone were investigated for the replacement of yeast extract. The structure of the lignocellulosic material was tackled by chemical treatment (1% NaOH) and enzymatic hydrolysis (Cellic® CTec2). B. coagulans DSM ID 14-300 was employed for the homofermentative conversion of the released hexose and pentose sugars to polymerizable L-(+)-LA of over 99.5% optical purity. The addition of autolyzed baker’s yeast led to the best results of fermentation, enabling an LA titer of 28.3 g L−1 and a yield of 91.6%.

Evaluation of alternative strategies for generating fermentable sugars from high-solids alkali pretreated sugarcane bagasse and successive valorization to L (+) lactic acid

Economical bioprocessing of lignocellulosic biomass essentially demands production of fermentable sugars in the concentrated form before their valorization. The present study aimed towards hydrolysis of alkali pretreated sugarcane bagasse at high solid loading and its successive valorization to L (+) lactic acid (LA). Two strategies were evaluated, wherein pretreatment, 12.5% substrate loading, Cellic CTec2 enzyme complex and thermophilic Bacillus coagulans NCIM 5648 were common to the processes. In Process A, when Cellic CTec2 was dosed at 30 FPU g−1 dry biomass, it hydrolyzed 75.8 ± 1.7% cellulose and 88.6 ± 2.1% xylan in 24 h. However, when its loading was changed to 25 mg protein g−1 glucan in Process B, Cellic CTec2 hydrolyzed 72.3 ± 0.3% and 68 ± 0.8% cellulose and xylan respectively. Valorization of glucose-rich filtrates obtained from Process A and B using two different media resulted in 50.4 ± 1.2 g L−1 and 51.24 ± 1.31 g L−1 of LA production from 54.7 to 62.7 g L−1 of glucose respectively. Attaining 1.75–2.4 g L−1 h−1 LA productivity with two scenarios of separate hydrolysis and fermentation is highly encouraging. It opens newer avenues for bio-based LA production using a greener approach.

Microwave processes: A viable technology for obtaining xylose from walnut shell to produce lactic acid by Bacillus coagulans

Biorefining uses alternative resources (such as biomass) instead of petrochemical sources to produce bioenergy and platform chemicals. Walnut shell is a promising lignocellulosic biomass for the production of fermentable carbohydrates. In this study, the platform chemical L-(+) lactic acid was produced after walnut shell had been treated with physicochemical and biological processes. So biomass waste can be regenerated to the benefit of the circular bio-economy. Autohydrolysis assisted by microwave processes was used to depolymerize hemicellulose and amorphous cellulose in walnut shell to yield 63.5% w/w of xylose in a liquid fraction (hydrolysate) with a minimum yield of glucose. The optimum reaction conditions to obtain a high xylose yield with a low byproduct titer were 190 °C for 25 min. L-(+) lactic acid was produced by thermophilic Bacillus coagulans DSM 2314 with a carbon source of xylose and glucose which were solubilized in hydrolysate from autohydrolyzed walnut shell. A non-sterile and straightforward fermentation medium coupled with a pH adjustment of the crude hydrolysate in an anaerobic environment led to a L- (+) lactic acid yield of 81% w/w with a productivity of 0.2 g/L/h.

Production and Purification of l-lactic Acid in Lab and Pilot Scales Using Sweet Sorghum Juice

Sweet sorghum juice (SSJ) was evaluated as fermentation substrate for the production of l-lactic acid. A thermophilic Bacillus coagulans isolate was selected for batch fermentations without the use of additional nutrients. The first batch of SSJ (Batch A) resulted on higher lactic acid concentration, yield and productivity with values of 78.75 g∙L−1, 0.78 g∙g−1 and 1.77 g∙L−1 h−1, respectively. Similar results were obtained when the process was transferred into the pilot scale (50 L), with corresponding values of 73 g∙L−1, 0.70 g∙g−1 and 1.47 g∙L−1 h−1. A complete downstream process scheme was developed in order to separate lactic acid from the fermentation components. Coarse and ultra-filtration were employed as preliminary separation steps. Mono- and bipolar electrodialysis, followed by chromatography and vacuum evaporation were subsequently carried out leading to a solution containing 905.8 g∙L−1 lactic acid, with an optical purity of 98.9%. The results of this study highlight the importance of the downstream process with respect to using SSJ for lactic acid production. The proposed downstream process constitutes a more environmentally benign approach to conventional precipitation methods.

L-Lactic acid production by Bacillus coagulans through simultaneous saccharification and fermentation of lignocellulosic corncob residue

l-Lactic acid is an important monomer of polylactic acid, a biodegradable plastic. In this study, we systematically investigated l-lactic acid fermentation by thermophilic Bacillus coagulans strains using the corncob residue (CCR), a widely available lignocellulosic material. Our results showed that 68.0 g/L l-lactic acid was obtained with a high yield of 0.85 g/g cellulose by B. coagulans strain H-1 after 36 h in batch fermentation. Strain H-1 could produce 79.1 g/L l-lactic acid with a yield of 0.76 g/g cellulose after 84 h in fed-batch fermentation. The results suggest that attractive lignocellulosic wastes such as CCR could be used in the efficient biomanufacture of l-lactic acid.

One step open fermentation for lactic acid production from inedible starchy biomass by thermophilic Bacillus coagulans IPE22

The aim of this study was to establish a simplified operational process for lactic acid (LA) production by Bacillus coagulans IPE22 from inedible starchy biomass with open fermentation method. First, 29.47 mU/mg specific amylase activity was detected in direct batch fermentation from soluble starch, but the activity of the produced amylase was too low for effective production of LA. Then seven batches from 72 g/L soluble starch were conducted without sterilization. It was found that one step simultaneous liquefaction, saccharification and fermentation (SLSF) with the addition of mesothermal α-amylase and glucoamylase was the optimal mode with LA concentration, yield and productivity of 68.72 g/L, 0.99 g/g and 1.72 g/L h respectively. Finally, inedible starchy biomass, cassava and sorghum flours, were proved to be alternatives to refined soluble starch. For the first time, one step open SLSF of inedible starchy biomass was reported for LA production by B. coagulans.

A Process Study of Lactic Acid Production from Phragmites australis Straw by a Thermophilic Bacillus coagulans Strain under Non-Sterilized Conditions

Phragmites australis straw (PAS) is an abundant and renewable wetland lignocellulose. Bacillus coagulans IPE22 is a robust thermophilic strain with pentose-utilizing capability and excellent resistance to growth inhibitors. This work is focused on the process study of lactic acid (LA) production from P. australis lignocellulose which has not been attempted previously. By virtue of thermophilic feature of strain IPE22, two fermentation processes (i.e., separated process and integrated process), were developed and compared under non-sterilized conditions. The integrated process combined dilute-acid pretreatment, hemicellulosic hydrolysates fermentation, and cellulose utilization. Sugars derived from hemicellulosic hydrolysates and cellulose enzymatic hydrolysis were efficiently fermented to LA in a single vessel. Using the integrated process, 41.06 g LA was produced from 100 g dry PAS. The established integrated process results in great savings in terms of time and labor, and the fermentation process under non-sterilized conditions is easy to scale up for economical production of lactic acid from PAS.

A novel α-galactosidase from the thermophilic probiotic Bacillus coagulans with remarkable protease-resistance and high hydrolytic activity.

A novel α-galactosidase of glycoside hydrolase family 36 was cloned from Bacillus coagulans, overexpressed in Escherichia coli, and characterized. The purified enzyme Aga-BC7050 was 85 kDa according to SDS-PAGE and 168 kDa according to gel filtration, indicating that its native structure is a dimer. With p-nitrophenyl-α-d- galactopyranoside (pNPGal) as the substrate, optimal temperature and pH were 55 °C and 6.0, respectively. At 60 °C for 30 min, it retained > 50% of its activity. It was stable at pH 5.0–10.0, and showed remarkable resistance to proteinase K, subtilisin A, α-chymotrypsin, and trypsin. Its activity was not inhibited by glucose, sucrose, xylose, or fructose, but was slightly inhibited at galactose concentrations up to 100 mM. Aga-BC7050 was highly active toward pNPGal, melibiose, raffinose, and stachyose. It completely hydrolyzed melibiose, raffinose, and stachyose in < 30 min. These characteristics suggest that Aga-BC7050 could be used in feed and food industries and sugar processing.

Non-sterilized fermentation of high optically pure d-lactic acid by a genetically modified thermophilic Bacillus coagulans strain

BackgroundOptically pure d-lactic acid (≥ 99%) is an important precursor of polylactic acid. However, there are relatively few studies on d-lactic acid fermentation compared with the extensive investigation of l-lactic acid production. Most lactic acid producers are mesophilic organisms. Optically pure d-lactic acid produced at high temperature not only could reduce the costs of sterilization but also could inhibit the growth of other bacteria, such as l-lactic acid producers.ResultsThermophilic Bacillus coagulans is an excellent producer of l-lactic acid with capable of growing at 50 °C. In our previous study, the roles of two l-lactic acid dehydrogenases have been demonstrated in B. coagulans DSM1. In this study, the function of another annotated possible l-lactate dehydrogenase gene (ldhL3) was verified to be leucine dehydrogenase with an activity of 0.16 units (μmol/min) per mg protein. Furthermore, the activity of native d-lactate dehydrogenase was too low to support efficient d-lactic acid production, even under the control of strong promoter. Finally, an engineered B. coagulans D-DSM1 strain with the capacity for efficient production of d-lactic acid was constructed by deletion of two l-lactate dehydrogenases genes (ldhL1 and ldhL2) and insertion of the d-lactate dehydrogenase gene (LdldhD) from Lactobacillus delbrueckii subsp. bulgaricus DSM 20081 at the position of ldhL1.ConclusionsThis genetically engineered strain produced only d-lactic acid under non-sterilized condition, and finally 145 g/L of d-lactic acid was produced with an optical purity of 99.9% and a high yield of 0.98 g/g. This is the highest optically pure d-lactic acid titer produced by a thermophilic strain.

Production of high concentration of l-lactic acid from oil palm empty fruit bunch by thermophilic Bacillus coagulans JI12.

Thermophilic Bacillus coagulans JI12 was used to ferment hemicellulose hydrolysate obtained by acid hydrolysis of oil palm empty fruit bunch at 50°C and pH 6, producing 105.4 g/L of l-lactic acid with a productivity of 9.3 g/L/H by fed-batch fermentation under unsterilized conditions. Simultaneous saccharification and fermentation (SSF) was performed at pH 5.5 and 50°C to convert both hemicellulose hydrolysate and cellulose-lignin complex in the presence of Cellic Ctec2 cellulases using yeast extract (20 g/L) as the nitrogen source, giving 114.0 g/L of l-lactic acid with a productivity of 5.7 g/L/H. The SSF was also conducted by replacing yeast extract with equal amount of dry Bakers' yeast, achieving 120.0 g/L of l-lactic acid with a productivity of 4.3 g/L/H. To the best of our knowledge, these lactic acid titers and productivities are the highest ever reported from lignocellulose hydrolysates.

Assessment of Compost Stability Using Single and Mixed Culture in a Static Semi-closed Fed-batch Reactor

Different microbial inoculums are expected to affect the degradation and stability of different compost materials. The results tend to vary more significantly in an open system due to the varied ambient conditions (temperature, moisture and air-borne microbial). To minimise such variation and avoid low reproducibility of the data, the present work was performed in a static semi-closed fed-batch reactor (SSCFBR) for better control of the composting process. The thermophilic Bacillus coagulans (BC) and the commercial effective microorganisms (EM) were used as the microbial inoculum (MI) in the SSCFBR. Distilled water was used in place of the MI for the control experiments. The mixture of chicken dung (CD), wooden husk (WH), and rice husk (RH) were used as the compost materials. The MI was inoculated into the compost bed in the SSCFBR (15 L working volume) at the same initial optical density (OD) of 0.8 and the initial volume of 800 mL with a forced aeration of 0.4 L/min. The parameters for the assessment of compost stability included the temperature, moisture content, oxygen uptake rate (OUR) and carbon dioxide emission rate (CER). BC showed comparable or better results to that achieved by the commercial EM. The temperature profile for both composts showed similar patterns where the highest peak of temperature was recorded within 1 d of composting (slightly higher for BC, 55 ± 0.8 °C and 47 ± 0.8 °C for EM). Such thermophilic temperature profile has not been observed in the control. The initial moisture content was set similar for all composts (44.5 ± 0.7 %). The moisture content declined slightly different for both MI compost (to 39.0 ± 0.6 % for BC and 36.6 ± 1.2 % for EM). Both composts showed very similar trend in the OUR. A higher evolution rate of carbon dioxide (CO2) was observed for the compost with BC for the first three days (0.06 ± 0.014 g/mol) as compared to that by EM (0.02 ± 0.007 g/mol). On the 4th d of composting, the production of CO2 in EM compost showed a near constant value (0.008 ± 0.002 g/mol) but the BC compost showed the rate of 0.06 ± 0.01 g/mol the 5th d and declined to 0.02 ± 0.004 g/mol on the 7th d. For both composting cases, the highest rate of CO2 was observed at the highest peak of temperature. The SSCFBR has been designed and could be used to facilitate the assessment of compost stability in a closed system with improved reproducibility of the data for composting.

Model-based characterisation of growth performance and l-lactic acid production with high optical purity by thermophilic Bacillus coagulans in a lignin-supplemented mixed substrate medium

Three Bacillus coagulans strains were characterised in terms of their ability to grow in lignin-containing fermentation media and to consume the lignocellulose-related sugars glucose, xylose, and arabinose. An optical-density high-throughput screening was used for precharacterisation by means of different mathematical models for comparison (Logistic, Gompertz, Baranyi, Richards & Stannard, and Schnute). The growth response was characterised by the maximum growth rate and lag time. For a comparison of the screening and fermentation results, an unstructured mathematical model was proposed to characterise the lactate production, bacterial growth and substrate consumption. The growth model was then applied to fermentation procedures using wheat straw hydrolysates. The results indicated that the unstructured growth model can be used to evaluate lactate producing fermentation. Under the experimental fermentation conditions, one strain showed the ability to tolerate a high lignin concentration (2.5g/L) but lacked the capacity for sufficient pentose uptake. The lactate yield of the strains that were able to consume all sugar fractions of glucose, xylose and arabinose was ∼83.4%. A photometric measurement at 280nm revealed a dynamic change in alkali-lignin concentrations during lactate producing fermentation. A test of decolourisation of vanillin, ferulic acid, and alkali-lignin samples also showed the decolourisation performance of the B. coagulans strains under study.

Cost-effective simultaneous saccharification and fermentation of l-lactic acid from bagasse sulfite pulp by Bacillus coagulans CC17

The main barriers to cost-effective lactic acid production from lignocellulose are the high cost of enzymes and the ineffective utilization of the xylose within the hydrolysate. In the present study, the thermophilic Bacillus coagulans strain CC17 was used for the simultaneous saccharification and fermentation (SSF) of bagasse sulfite pulp (BSP) to produce l-lactic acid. Unexpectedly, SSF by CC17 required approximately 33.33% less fungal cellulase than did separate hydrolysis and fermentation (SHF). More interestingly, CC17 can co-ferment cellobiose and xylose without any exogenous β-glucosidase in SSF. Moreover, adding xylanase could increase the concentration of lactic acid produced via SSF. Up to 110g/L of l-lactic acid was obtained using fed-batch SSF, resulting in a lactic acid yield of 0.72g/g cellulose. These results suggest that SSF using CC17 has a remarkable advantage over SHF and that a potentially low-cost and highly-efficient fermentation process can be established using this protocol.

Lactic acid production in a membrane bioreactor system with thermophilic Bacillus coagulans: Online monitoring and process control using an optical sensor

ABSTRACTMembrane bioreactor systems can efficiently reduce product inhibition in lactic acid production processes, enhancing their productivity. Filtration parameters, e.g. crossflow velocity and transmembrane pressure, were controlled with varying biomass concentration, which was monitored using an optical sensor in real time. The filtration flux was successfully maintained as desired according to the control strategy. We also investigated the effect of feed concentration and dilution rate on process characteristics. The continuous fermentation remained stable for more than 80 h, achieving the highest productivity of 8.6 g·L–1·h–1 with a feed concentration of 70 g·L–1 and a dilution rate of 0.2 h–1.

Highly efficient production of optically pure l-lactic acid from corn stover hydrolysate by thermophilic Bacillus coagulans

A thermophilic strain Bacillus coagulans (NBRC 12714) was employed to produce l-lactic acid from corn stover hydrolysate in membrane integrated continuous fermentation. The strain NBRC 12714 metabolized glucose and xylose by the Embden–Meyerhof-Parnas pathway (EMP) and the pentose phosphate pathway (PPP), producing l-lactic acid with optical purity >99.5%. The overall l-lactic acid titer of 92g/l with a yield of 0.91g/g and a productivity of 13.8g/l/h were achieved at a dilution rate of 0.15h−1. The productivity obtained was 1.6-fold than that of conventional continuous fermentation without cell recycling, and also was the highest among the relevant studies ever reported. These results indicated that the process developed had great potential for economical industrial production of l-lactic acid from lignocellulosic biomass.