Steeping Process Research Articles

Effects of thermophilic and acidophilic microbial consortia on maize wet-milling steeping

To understand the ecology of species and promote biotechnology through beneficial strain selection for improving starch yield in maize wet-milling steeping, bacterial diversity and community structure during the counter-current steeping process in a commercial steeping system were characterized and investigated. The microbial diversity in the steeping liquor, which consisted of 16 phyla, 131 families, and 290 genera, was more abundant compared to those present on the surface of unsteeped maize. As the counter-current steeping progressed, exposing newer maize to the older steepwater, Lactobacillus dominated, replacing Rahnella, Pseudomonas, Pantoea, and Serratia. The thermophilic and acidophilic microbial consortia were enriched through adaptive evolution engineering and employed to improve starch yield. Several steeping strategies were evaluated, including water alone, SO2 alone, mono-culture of B. coagulans, microbial consortia, and a combination of consortium and SO2. Combining the microbial consortium with SO2 significantly increased the starch yield to, about 66.4 ± 0.5%, a 22% and 46% increase over SO2 alone and the consortium alone, respectively. Scanning electron microscope (SEM) of steeped maize structure indicated that the combination of consortium and SO2 disrupted the protein matrix and widened gaps between starch granules in maize endosperm. This released proteins into the steepwater and left starch granules in the aleurone layer. The steeping strategy of using thermophilic and acidophilic microbial consortium as additives shows potential application as an environmentally friendly alternative to conventional maize steeping procedures.

Modification of starch digestibility and phenolic bioaccessibility in phenolic-rich flours by physical treatments with ferulic acid followed by vacuum oven and freeze-drying

Consumer demand for healthy snack products has led to increased efforts to modify glycemic responses of these products. The formation of starch-ferulic acid (FA) complexes through acidified water steeping leads to an increase in RS content and reduction of glycemic responses in animal models. However, the application of these methodologies to commercially relevant phenolic-rich flours (black rice, BR; purple maize, PM) and their impacts on starch digestibility and phenolic bioaccessbility remain unclear. Using an acidified water steeping method previously reported in pure starch, potato starch (PS), BR and PM flour slurries were prepared with FA at a 20:1 and 8:1 ratio in HCl-acidified water (pH 2). The steeping process reduced phenolic content (∼75%) and increased starch digestibility. Ferulic acid loading capacity on PS (22–124 mg/g starch) was greater than for PM and BR flours (8–40 mg/g starch). Ferulic acid bioaccessibility decreased with an increase in its amount in PS and flours. Steeping enhances cyanidin 3-O-glucoside bioaccessibility, while addition of FA decreases its bioaccessibility. The increase in starch digestibility of phenolic-rich flours after steeping with and without FA can be attributed to the limited loading capacity of FA and loss of native phenolics during the steeping process.

Brewing Excellence: Improving Coffee Quality with the V60 Stirred-agitation Technique

Manual coffee brewing is the preferred method owing to its capacity to produce a more delicate and diverse range of flavors. While extensive research has been conducted on the process of coffee brewing, there has been limited discourse on the impact of agitation techniques on the quality of the steeping phase. The primary objective of this study is to explore the influence of agitation methods, specifically V60 and Kalita, on the quality of the steeping process. A survey-based investigation was conducted, analyzing sensory attributes such as flavor, aftertaste, acidity, body, balance, sweetness, overall flavor, acid intensity, and body level. A panel of twenty skilled baristas participated as evaluators. The findings of this research demonstrate that agitation plays a crucial role in achieving optimal extraction during the coffee brewing procedure. The particular agitation method employed distinguishes the brewing process and the final steeping outcome. Notably, the use of stirred agitation in the V60 method results in a well-balanced steeping flavor characterized by an acidic orange aroma and a subtle undertone of brown sugar, all without leaving a lingering, cloying aftertaste. The brewed coffee produced using this method exhibits a moderate level of acidity and body, alongside a subtly sweet flavor profile that contributes to an enhanced overall taste experience.

Purification of lipopeptide biosurfactant extracts obtained from a complex residual food stream using Tricine-SDS-PAGE electrophoresis.

Protocols to identify lipopeptide biosurfactant extracts contained in complex residual streams are very important, as fermented agri-food matrices are potential sources of these valuable compounds. For instance, corn steep liquor (CSL), a secondary stream of the corn wet-milling industry, is composed of a mixture of microbial metabolites, produced during the corn steeping process, and other natural metabolites released from corn, that can interfere with the purification and analysis of lipopeptides. Electrophoresis could be an interesting technique for the purification and further characterization of lipopeptide biosurfactant extracts contained in secondary residual streams like CSL, but there is little existing literature about it. It is necessary to consider that lipopeptide biosurfactants, like Surfactin, usually are substances that are poorly soluble in water at acidic or neutral pH, forming micelles what can inhibit their separation by electrophoresis. In this work, two lipopeptide biosurfactant extracts obtained directly from CSL, after liquid-liquid extraction with chloroform or ethyl acetate, were purified by applying a second liquid extraction with ethanol. Following that, ethanolic biosurfactant extracts were subjected to electrophoresis under different conditions. Lipopeptides on Tricine-SDS-PAGE (polyacrylamide gels) were better visualized and identified by fluorescence using SYPRO Ruby dye than using Coomassie blue dye. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis of lipopeptide isoforms separated by electrophoresis revealed the presence of masses at 1,044, 1,058, and 1,074m/z, concluding that Tricine-SDS-PAGE electrophoresis combined with MALDI-TOF-MS could be a useful tool for purifying and identifying lipopeptides in complex matrices.

Effect of a Biosurfactant Extract Obtained from a Corn Kernel Fermented Stream on the Sensory Colour Properties of Apple and Orange Juices.

In this work, we assessed the effect of a biosurfactant extract, which possesses preservative properties, on the sensory properties, regarding colour, of two fruit juices: pasteurized apple juice and natural orange juice. This biosurfactant extract was obtained from corn steep liquor, a secondary stream of the corn wet-milling industry. The biosurfactant extract is composed of natural polymers and biocompounds released during the spontaneous fermentation of corn kernels during the steeping process of corn. The reason for this study is based on the importance of colour as a visual attribute that can determine the consumer's preferences; it is important to study the effect of the biosurfactant extract under evaluation before including it in juice matrices. For this, a surface response factorial design was employed and the effects of the biosurfactant extract concentration (0-1 g/L), the storage time (1-7 days), and the conservation temperature (4-36 °C) on the CIELAB colour parameters (L*, a*, b*) of the juice matrices were evaluated, as well as the total colour differences (ΔE*) regarding the control juices and the saturation index (Cab*). Moreover, the CIELAB coordinates of each conducted treatment were converted into RGB values to obtain visual colour differences that can be appreciated by testers or consumers.

Metaheuristic Algorithms to Optimal Parameters Estimation of a Model of Two-Stage Anaerobic Digestion of Corn Steep Liquor

Anaerobic Digestion (AD) of wastewater for hydrogen production is a promising technology resulting in the generation of value-added products and the reduction of the organic load of wastewater. The Two-Stage Anaerobic Digestion (TSAD) has several advantages over the conventional single-stage process due to the ability to control the acidification phase in the first bioreactor, preventing the overloading and/or the inhibition of the methanogenic population in the second bioreactor. To carry out any process research and process optimization, adequate mathematical models are required. To the best of our knowledge, no mathematical models of TSAD have been published in the literature so far. Therefore, the authors’ motivation is to present a high-quality model of the TSAD corn steeping process for the sequential production of H2 and CH4 considered in this paper. Four metaheuristics, namely Genetic Algorithm (GA), Firefly Algorithm (FA), Cuckoo Search Algorithm (CS), and Coyote Optimization Algorithm (COA), have been adapted and implemented for the first time for parameter identification of a new nonlinear mathematical model of TSAD of corn steep liquor proposed here. The superiority of some of the algorithms has been confirmed by a comparison of the observed numerical results, graphical results, and statistical analysis. The simulation results show that the four metaheuristics have achieved similar results in modelling the process dynamics in the first bioreactor. In the case of modelling the second bioreactor, a better description of the process dynamics trend has been obtained by FA, although GA has acquired the lowest value of the objective function.

Variation in Volatile Compounds of Raw Pu-Erh Tea upon Steeping Process by Gas Chromatography-Ion Mobility Spectrometry and Characterization of the Aroma-Active Compounds in Tea Infusion Using Gas Chromatography-Olfactometry-Mass Spectrometry.

Steeping process is an important factor for aroma release of tea, which has rarely been investigated for the aroma changes of raw Pu-erh tea (RAPT). In addition, the comprehensive aroma characteristics identification of RAPT infusion is necessary. In this study, GC-IMS coupled with principal component analysis (PCA) was used to clarify the difference of volatile profiles during the steeping process of RAPT. Furthermore, the volatiles contained in the RAPT infusion were extracted by three pretreatment methods (HS-SPME, SBSE, and SAFE) and identified using GC-O-MS. According to the odor activity value, 28 of 66 compounds were categorized as aroma-active compounds. Aroma recombination and omission experiments showed that "fatty", "green", "fruity", and "floral" are considered to be the main aroma attributes of RAPT infusion with a strong relationship with 1-octen-3-one, 1-octen-3-ol, (E)-2-octenal, β-ionone, linalool, etc. This study will contribute a better understanding of the mechanism of the RAPT steeping process and volatile generation.

Inhibition of Chitosan with Different Molecular Weights on Barley-Borne Fusarium graminearum during Barley Malting Process for Improving Malt Quality.

There are many Fusarium graminearum contaminations in barley that are often associated with malt and beer quality issues. Thus, it is important to find a biological antifungal agent to prevent the growth of F. graminearum during malting. Minimum inhibition concentration (MIC) of chitosan for mycelial growth and spore germination of F. graminearum was 2.6 g/L and 1.6 g/L, respectively, indicating that the F. graminearum strain was highly sensitive toward chitosan. Chitosan with a molecular weight of 102.7 kDa was added at 0.5 g/kg during the first steeping stage, resulting in the maximum inhibition rate of F. graminearm in barley. The biomass of F. graminearm and deoxynivalenol content in the infected barley at the end of germination with 0.5 g/kg chitosan treatment were decreased by 50.7% and 70.5%, respectively, when compared with the infected barley without chitosan. Chitosan could remove the negative effects of F. graminearm infection on barley germination and malt quality, which makes the application of chitosan during the steeping process as a potential antifungal agent in the malting process to protect from F. graminearum infection.

Optimisation of Malting Parameters for Quinoa and Barley: Application of Response Surface Methodology

Quinoa (Chenopodium quinoa Willd) is a nutritious pseudocereal that is more stress-tolerant compared with traditional cereals. It is an excellent example of a climate-smart crop that is more resilient to climate change compared with barley. The purpose of the study was to investigate the optimum malting conditions required to produce quinoa malt using barley as a control. Response surface methodology (RSM) was used to investigate the influence of the two malting parameters steeping time and germination time on Brix (wort extract), diastatic power (DP), and free amino nitrogen (FAN) of the malt. The temperature was set at 15°C during the steeping process. Steeping time ranging from 12 to 48 hours and germination time ranging from 24 to 96 hours were designed using a central composite design (CCD). The kilning temperature for all malts was 65°C. For quinoa malt, there was a notable weak positive correlation between germination time and Brix (r = +0.119). However, there was a strong positive correlation between steeping time and diastatic power (r = +0.893). A similar trend was noted for barley with a weak positive correlation between germination time and Brix (r = +0.142). A strong positive correlation was also recorded between steeping time and diastatic power (r = +0.897) during the malting of barley. There was a relatively stronger correlation between steeping time and FAN (r = +0.895) than germination time and FAN (r = +0.275) in quinoa malt. The optimum values for the malting of barley were 47.68 hrs steeping time and 82.55 hrs germination time with a desirability value of 1.00. The responses for the optimised barley malt were 8.25°Bx, 162.28 mg/L, and 271.69°L for Brix, FAN, and diastatic power, respectively. To produce quinoa malt with Brix, FAN, and diastatic power of 8.37°Bx, 165.60 mg/L, and 275.86°L, respectively, malting conditions of 47.69 hrs steeping time and 95.81 hrs germination time are required. It was noted that quinoa is a very good candidate for producing high-quality malt for the brewing process.

Evaluation of Calcium Alginate-Based Biopolymers as Potential Component of Membranes for Recovering Biosurfactants from Corn Steep Water

Corn steep water (CSW) is a complex agro-food stream that is used as a source of cost-competitive biosurfactants, since they are produced spontaneously in the steeping process of corn, avoiding production costs. Nevertheless, the extraction of biosurfactants from CSW using sustainable processes is still a challenge. Consequently, the use of calcium alginate membranes could present a novel and sustainable technology for recovering biosurfactants from aqueous streams. Therefore, the aim of this work is to evaluate calcium alginate-based biopolymers, without and with the presence of grape marc as an additive, as a key component of membranes for the recovery of biosurfactants in corn steep water. Biosurfactants are present in CSW, together with other inorganic solutes and biomolecules, such as organic acids, sugars, cations, anions as well as metals. Hence, the competition of these mentioned compounds for the active sites of the calcium alginate-based biopolymers was high. However, they showed a good adsorption capacity for biosurfactants, recovering around 55 ± 2% and 47 ± 1%, of biosurfactants from CSW using both calcium alginate-based biopolymers, with and without biodegraded grape marc. Regarding adsorption capacity, it was 54.8 ± 0.6 mg biosurfactant/g bioadsorbent for the biopolymer containing grape marc, and 46.8 ± 0.4 mg biosurfactant/g bioadsorbent for the calcium alginate-based biopolymer alone. Based on these results, it could be postulated that the formulation of green membranes, based on calcium alginate-based polymers, could be an interesting alternative for the recovery of biosurfactants from aqueous streams including CSW.

TAHIL VE PSEUDO-TAHILLARIN ISLAK ÖĞÜTME TEKNOLOJİSİ

Öğütmenin amacı; tohumu meydana getiren başlıca kısımları, öğütme yöntemine göre ayırarak, gıda endüstrisi veya diğer endüstrilerde kullanmaktır. Öğütme, genel olarak kuru ve ıslak öğütme olarak sınıflandırılabilir. Islak öğütmede amaç; nişasta, protein, besinsel lif ve ham yağ gibi tohumun başlıca kimyasal bileşenlerini ayırmak iken; kuru öğütmede amaç tanenin anatomik kısımları olan endosperm, ruşeym ve kepeği ayırmaktır. Islak öğütmede her bileşen mümkün olan en saf haliyle ayrılırken; kuru öğütmede, yüksek kalitede rafine veya tam tane unu elde edilir. Islak öğütme, tahıl veya pseudo-tahılların temel bileşenlerini fiziksel, kimyasal, biyokimyasal ve mekanik işlemler ile ayıran endüstriyel bir işlemdir. Islak öğütme, ıslatma işlemi ile başlar, ardından mekanik ayırma işlemi gelir. Islak öğütme işlemi sanayide, çoğunlukla buğday ve mısıra uygulanırken, sorgum, arpa, yulaf ve pirinç gibi tahıllara da uygulanmaktadır. Karabuğday, kinoa ve amarant pseudo-tahıllarının ıslak öğütmesi ile ilgili çalışmalar henüz laboratuvar ölçeğindedir. Tahıl veya pseudo-tahılların ıslak öğütme koşulları tamamen nişasta verimini ve fizikokimyasal özelliklerini etkilemektedir.

Prediction of suitable brewing cuppages of Dahongpao tea based on chemical composition, liquor colour and sensory quality in different brewing

Oolong tea is famous for its characteristic of durably brewing. To explore suitable brewing cuppages and the scientific methods to brew Oolong tea in multiple steeping process. Dahongpao tea (Zhengyan, Banyan and Zhouyan tea) is well known Oolong tea variety, brewed at 14 times and assessed its chemical composition, infusion colour and sensory quality in different brewing intervals. The results showed that Zhengyan tea (A3) had the best quality of steeping among the chosen tea. It could be brewed up to 10 cuppages with 80% sensory score. The chemical composition and tea infusion colour strength were higher in Zhengyan tea. Though, 70% caffeine leached within first three steeping. The Forest regression model revealed that the suitable brewing time ranges between 4 and 10 in the chosen Dahongpao tea variety. This study provides a scientific method and suitable steeping times for the drinking of different Dahongpao tea through dynamic analysis of quantity of chemical composition, infusion colour strength and sensory quality.

Characterization of arabinoxylan degrading enzymes during wheat malting

In this paper the arabinoxylanase activities and the content of water-extractable arabinoxylans (WEAXs) including their degradation products in wheat Yannong 24 during the malting process were studied. The results showed that the endo-β-1,4-xylanase activity had a large increase during the steeping process, endo-β-1,4-xylanase and α-L-arabinofuranosidase achieved the highest activity on day 4 of germination. During malt kilning, endo-β-1,4-xylanase and α-L- arabinofuranosidase were not heat stable, and the activity decline rate were 65.88% and 69.58%; whereas malt β-D-xylosidase was resistant to heat and it’s activity decline rate was only 18.74%. It is during steeping and the first day of germination that the content of WEAXs and it’s degradation products (WEAXs+DP) dramatically increased, and this corresponded to the greater improvement of endo-β-1,4-xylanase activity in the same phase. The WEAXs+DP content has a very significant positive correlation with the endo-β-1,4-xylanase activity (0.892, p<0.01). It can be indicated that endo-β-1,4-xylanase is the main degrading enzyme in the degradation of WUAXs and the increase of the WEAXs content during steeping and the first day of wheat germination.

Plastic Teabags Release Billions of Microparticles and Nanoparticles into Tea.

The increasing presence of micro- and nano-sized plastics in the environment and food chain is of growing concern. Although mindful consumers are promoting the reduction of single-use plastics, some manufacturers are creating new plastic packaging to replace traditional paper uses, such as plastic teabags. The objective of this study was to determine whether plastic teabags could release microplastics and/or nanoplastics during a typical steeping process. We show that steeping a single plastic teabag at brewing temperature (95 °C) releases approximately 11.6 billion microplastics and 3.1 billion nanoplastics into a single cup of the beverage. The composition of the released particles is matched to the original teabags (nylon and polyethylene terephthalate) using Fourier-transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). The levels of nylon and polyethylene terephthalate particles released from the teabag packaging are several orders of magnitude higher than plastic loads previously reported in other foods. An initial acute invertebrate toxicity assessment shows that exposure to only the particles released from the teabags caused dose-dependent behavioral and developmental effects.

Centrifuge modelling of submarine landslides due to static liquefaction

Sand erosion and scouring caused by waves and marine currents result in gradual increase of local seabed inclination and formation of slopes around hydraulic structures and offshore foundations. During this process, shear stresses in the soil body increase monotonically which may lead to static liquefaction and damage of the adjacent offshore infrastructure. This paper presents the details of a newly developed static liquefaction triggering actuator to be used at an enhanced gravity condition in a geotechnical centrifuge. This actuator simulates the steeping process of submarine sand layers due to scouring and enables the investigation of failure mechanisms in submerged slopes. The details of the centrifuge test set-up designed and constructed to simulate the process of triggering static liquefaction in loose sand layers are presented. Furthermore, the performance of the novel integrated model preparation facility using sand fluidization is explained. The set-up was used to conduct several centrifuge tests at four different slope steepening rates to investigate the slope steepening rate effects. Moreover, the effect of viscosity of the submerging pore fluid on the behaviour of the slopes at the onset of failure is investigated. The Coriolis effect on loose saturated sand samples during increase of g-level is examined as well. Results show that the built-up of pore pressure due to local shear deformations can be detected and considered as one of the triggering mechanisms of this kind of submarine slope instabilities.

Päťdesiat rokov hurbanovskej sladovne

In April 2017 it was exactly half a century since construction work on malting plant was completed, and in October it was 50 years since the first tonnes of Hurbanovo malt had been produced. The maltery was then a part of the state-owned enterprise Zapadoslovenske pivovary a sladovne, Bratislava. Once everything was completed, the maltery consisted of 64 steeping tanks, 16 Wanderhaufen system lines and four double-floor kilns. The planned malt production capacity of 30,000 tonnes was already achieved in 1968. Organisational change in 1984 caused the creation of the separate state-owned enterprise Zlatý Bažant, Hurbanovo. After the new germination facility with eight Saladin boxes and two single-floor kilns were built and at the same time the steeping process was adjusted, the annual production capacity of maltery increased during 1986 and 1987 to approximately 63,000 tonnes of malt. In 1992, the brewery was transformed into a joint-stock company Zlatý Bažant and in 1995, the whole plant became a part of the Dutch company Heineken. In 1998, a new steeping room with nine cylindroconical steeping tanks took the place of the old steep tank system. Following adjustments of the technological process enabled the increase of total malt production capacity to 80,000 tonnes annually. Another organisational change happened in 2001, when maltery became an independent joint-stock company under the name Heineken Slovensko Sladovne. In 2002, there was a modernization, an increase in the capacity of six steeping tanks, a construction of six new circular germinating boxes and high-capacity single-floor kiln, thus extending the maximum annual production capacity to 150,000 tonnes. At the same time, the malt production in the outdated Wanderhaufen system floors and four original double-floor kilns was discontinued. The old Saladin boxes and two old singlefloor kilns were decommissioned gradually during 2012 and 2013. With an annual malt production capacity of around 111,000 tonnes, the maltery is the largest not only in Slovakia, but also in the entire Central Europe.

Fate of zearalenone, deoxynivalenol and deoxynivalenol-3-glucoside during malting process

Malting was simulated using two different batches of barley as raw material: a naturally contaminated batch and laboratory inoculated (with a deoxynivalenol (DON) and zearalenone (ZEA) producing Fusarium graminearum strain) one. Up to three contamination levels were prepared, every process being carried out in triplicate. A significant washout effect on DON was observed by the end of the first water phase (between 22.4 and 34% reduction) with an even more pronounced reduction (up to 75% decrease) by the end of the steeping process. ZEA content remained almost unchanged (no significant difference between the initial and the final concentration). Germination was characterized by an increase in all the three toxins (ZEA, DON and DON-3-G) concentrations, however showing a decreasing trend in the last 24 h of the stage, compared to the first day of germination. Kilning lead to a significant reduction of DON in the naturally contaminated batch (46.6 and 78.8%), nevertheless an increase in all other toxins and contamination levels was observed.

Effect of intermittent high-intensity sonication and temperature on barley steeping for malt production

Barley malt production comprises three main steps: steeping, germination, and drying. Ultrasound technology has been widely studied to find ways to improve mass transfer in food processes and, consequently, to reduce process times. So, this study evaluated the effect of temperature and the intermittent application of ultrasound on the steps involved in barley hydration. The barley hydration was carried out at 10, 15, 20, and 25 °C with and without the application of 0.75 W/mL and 1.5 W/mL of nominal power density at 20 kHz. The ultrasonic energy delivered was measured in the same conditions as the steeping process using a calorimetric method, taking distinct differential volume measurements throughout the hydration medium. The ultrasonic energy delivered presented average values of 51.1 W at 750 W and 84.7 W at 1500 W nominal power. Ultrasound application increased both water uptake rates and equilibrium moisture content as shown by the Peleg and Weibull-exponential model parameters, with the latter showing better adjustment (Radj2>0.953 and NRMSE<5%). Applying ultrasound also significantly reduced the time required to achieve the conventional moisture level required for barley germination: 29% and 44% at controlled temperatures of 20 °C and 25 °C, respectively.

Enhancing the production of phenolic compounds during barley germination by using chitooligosaccharides to improve the antioxidant capacity of malt.

To enhance the production of phenolic compounds during barley germination using chitooligosaccharide as an elicitor to improve the antioxidant capacity of malt. When used as an elicitor for barley germination, chitooligosaccharide with a molecular weight of 3kDa, added at 10mg/kg barley kernels during the first steeping cycle, led to the maximum production of phenolic compounds. Compared with the control with no chitooligosaccharide added to the steeping water, the total phenolic content was increased by 54.8%. Increases in the total phenolic content of the barley malt occurred when chitooligosaccharide was applied during the first or both the first and the second steeping cycles. Thus the antioxidant capacity of barley malt was increased significantly by adding chitooligosaccharide during the steeping process. Applying chitooligosaccharides during the steeping process increased the content of phenolic compounds thus improving the antioxidant capacity of the barley malt.

Immunostimulatory and anti-metastatic activity of polysaccharides isolated from byproducts of the corn starch industry

Corn steep liquor (CSL) is a major by-product of the corn steeping process that is utilized in the wet milling industry. To develop new physiologically active polysaccharides from CSL, polysaccharides were isolated and their innate immunostimulatory and anti-metastatic activities were investigated. Corn byproduct polysaccharides (CBP) were preferentially isolated from CSL and further separated into supernatant (CBP1S) and precipitate (CBP1P) fractions. The anti-complementary activity of CBP1S was more potent than CBP1P and CBP. In addition, CBP1S enhanced production of macrophage-stimulating cytokines (e.g., IL-6 and IL-12) and natural killer (NK) cell-activating substances (e.g., granzyme and interferon-γ). Further, CBP1S significantly inhibited lung metastasis at a dose of 1000μg per mouse in an experimental lung metastasis model. These results suggest that CBP1S seems to promote the inhibition of lung metastasis through a mechanism leading to stimulation of the innate immune system, and CBP1S could be used as immunostimulating agents and for industrial applications.